Characterization and epitope mapping of a new panel of monoclonal antibodies to estradiol receptor.
A new panel of monoclonal antibodies to the calf uterus estrogen receptor was prepared. Thirteen antibodies were characterized for their isotype and for the affinity for the antigen. These antibodies recognize the human receptor and can be used in Western blot analysis. The location of the epitopes was mapped on the antigen structure using synthetic fragments of estrogen receptor, and it was possible to group the antibodies in five groups. Many antibodies were useful for the purification of estrogen receptor from tissue extracts by immunoaffinity chromatography. The reciprocal inhibition of the antibodies for the antigen binding was measured with an immunoadsorption assay. This was maximal and symmetrical for antibody pairs within the same group, but was incomplete and, in some instances, asymmetrical between pairs of antibodies from different groups. One antibody was able to inhibit the estrogen receptor-DNA interaction, whereas two others were unable to recognize the receptor-DNA complexes. This new panel of antibodies is a useful addition to the existing tools for studying structure and function of the estrogen receptor.


Biomass yields of Chlorella from iron (Yx/Fe) in iron-limited batch cultures.
The maximum biomass in iron-limited photosynthetic batch cultures of chlorella increased as the logarithm of the iron concentration. The growth yield from iron (UxFe) showed a marked inverse relation to the specific growth rate. The maximum biomass yield, g dry biomass/g iron consumed, was 7.5 X 10(3) with specific growth rate 0.108h-1; the minimum was 0.79 X 10(3) with specific growth rate 0.145 h-1. The maximum specific growth rate in the exponential phase of Fe limited cultures varied as the initial Fe concentration. Fe-limited growth made the cells adhere to a glass surface.


Spatial and temporal expression profile of sex steroid receptors and antioxidant enzymes in the maternal-fetal interface of domestic cats.
Sex steroids and antioxidant enzymes modulate uterine and placental physiology. Failures in the expression, signaling, and/or secretion of these mediators are associated with female infertility and gestational problems. However, there is no data on the expression profile of receptors for sex steroids and antioxidant enzymes in the maternal-fetal interface of domestic cats. Uterus and placenta samples from non-pregnant diestrus cats and cats in mid- and late pregnancy were used to analyze the protein and gene expression of the receptors for estrogen alpha (ERα), progesterone (PR), and androgen (AR) and the antioxidant enzymes superoxide dismutase 1 (SOD1), catalase, and glutathione peroxidase 1 (GPX1) by immunohistochemistry and qPCR. Higher uterine expression of ERα, Pr, and Sod1 was observed in the pregnant cats, especially in mid-pregnancy, compared to non-pregnant diestrus cats, as well as reduced endometrial catalase immunostaining. In the placenta, the mRNA expression of Erα, Pr, Ar, and Gpx1 was higher in late pregnancy in relation to mid-pregnancy. Moreover, weak or no placental expression was observed for catalase in mid- and late pregnancy, while strong immunostaining was observed for AR in trophoblasts and decidual cells in mid-pregnancy. The findings of this study demonstrated that pregnancy in female cats increases the uterine expression of sex steroid receptors and antioxidant enzymes, and that the placental expression of these mediators varies according to gestational age.


Structure of the rigid-layer of Rhizobium cell wall. III. Electron microscopic evidence for the cellulose microfibrils association with peptidoglycan sacculi.
The morphology of peptidoglycan layer of Rhizobium cell wall was examined by transmission electron microscopy. Peptidoglycans were isolated from intact cells after treatment with sodium dodecyl sulfate, extraction with aqueous 45% phenol and then with a mixture of chloroform-methanol. Finally rigid layers were digested with trypsin and chymotrypsin. The results indicate the presence of lump or bar-like structures on the surface of the cell shaped peptidoglycan sacculi. Evidence is provided suggesting that the cellulose microfibrils arise directly from these excrescences found on the peptidoglycan surface. Digestion with cellulase removed all cellulose microfibrils whereas the lumps and bars remained as an integral part of the Rhizobium peptidoglycan.


Rare non-papillary lithiasis of calcium oxalate monohydrate generated on a central core of potassium urate.
This report describes a patient who developed a spheroidal calculus with a central part composed of potassium urate, surrounded by a continuous layer of calcium oxalate monohydrate with crystals of calcium oxalate dihydrate on the surface. The mechanism of calculus development is also suggested.


Limitations of pesticide genotoxicity testing using the bacterial in vitro method.
The results of genotoxicity assessment of 115 technical grade pesticide products (61 names) with the Ames test and mammalian erythrocyte micronucleus test in vivo are presented. In most cases, unidirectional results were found. However, some pesticides belonging to classes of neonicotinoids, derivatives of phenylurea, glycine and phenoxycarboxylic acids showed genotoxic effects only under in vivo conditions. All observed effects were statistically significant and dose-depended but were only slightly beyond the upper limit of the historical negative control. Pendimethalin (dinitroaniline pesticide) showed positive results only in Ames test. Furthermore, the limitations of Ames test related to the high cytotoxicity of pesticides against bacterial cells were shown (e.g. bipyridylium derivatives, chlorothalonils, dithiocarbamates, quinones, phenyl pyridinamines and sulphonylureas). But many pesticides possessing the highest cytotoxic activity against bacterial cells have shown low toxicity in vivo (MTD > 2000 mg/kg b.w.). On the other hand, all of the tested neonicotinoids, organophosphates, pyrethroids, phenoxycarboxylic acid derivatives were highly toxic to mice but non-cytotoxic against microbial cells. Triazole pesticides exerted toxicity to the target organ (bone marrow). Therefore, these findings suggest that genotoxicity assessment using at least two methods is needed for reliable evidence of the safe use of pesticides.


Experience of the first 1127 COVID-19 Vaccine Allergy Safety patients in Hong Kong - Clinical outcomes, barriers to vaccination, and urgency for reform.
Hong Kong has had a low incidence of COVID-19 vaccine related anaphylaxis, partly due to its Vaccine Allergy Safety (VAS) guidelines for screening those at higher risk of COVID-19 vaccine-associated allergic reactions. We characterize the initial experience of the VAS clinics, as well as the impact of unnecessary referrals to the vaccination program. All patients attending the VAS Clinics of the public and private health services between February and June 2021 were reviewed. Out of 1127 patients assessed at VAS clinics, 1102 (97.8%) patients were recommended for vaccination. Out of those contacted, more than 80% (450/558) received vaccination successfully; the remaining had not yet booked their vaccinations. The majority (87.5%) of patients not recommended was due to potential excipient allergies. Males were significantly more likely to be recommended (OR = 5.822, 95% CI = 1.361-24.903, p = 0.007), but no other features were associated with recommendation for vaccination. Almost half (45.1%) of public service referrals were rejected due to insufficient information or incorrect indications for referral. The majority of cases (56.2%) of patients referred for suspected "anaphylaxis" did not fulfil diagnostic criteria. COVID-19 vaccination is very safe and 98% of high-risk patients were recommended for vaccination. Barriers to VAS include a high proportion of inappropriate referrals, inaccurate diagnoses of anaphylaxis and inability to diagnose excipient allergies. Our data validates that a prior history of COVID-vaccine unrelated anaphylaxis should be removed as a precaution for vaccination. Closer collaborations between primary care and allergy specialists and changes in pharmaceutical legislation should be made a priority to promote vaccination uptake.


Natural radioactivity and radon exhalation rates in man-made tiles used as building materials in Japan.
Man-made tiles frequently used in Japan were collected, and activity concentrations and radon ((222)Rn) exhalation rates in these tiles were measured. Dose estimations for inhabitants living in houses built using these tiles were also carried out. The activity concentrations of (226)Ra, (228)Ra and (40)K in the man-made tiles were 31-170, 35-110 and 260-980 Bq kg(-1), respectively. The (222)Rn exhalation rates in the tiles were 8.8-21 μBq m(-2) s(-1). The ranges of experimental activity concentrations and (222)Rn exhalation rates were almost identical to those of natural rocks used as typical building materials in Japan. The maximum value of effective dose to inhabitants living in houses built with the man-made tiles was 0.14 mSv y(-1), which is lower than the reference level range (1-20 mSv y(-1)) for abnormally high levels of natural background radiation published in the ICRP Publication 103.


Death in the CNS: six-microns-under.
During embryonic development, large numbers of apoptotic cells are rapidly cleared by phagocytes. In this issue, Kurant et al. (2008) describe a new phagocytic receptor, called six-microns-under (SIMU), that promotes engulfment of apoptotic neurons by glial cells in the developing nervous system of Drosophila.


Modified gloves: A chance for the prevention of nosocomial infections.
Non-sterile gloves primarily serve as a barrier protection for health care workers (HCWs). However, pathogens may often contaminate the skin of HCWs during glove removal; therefore, pathogens may be further transmitted and cause nosocomial infections. A field study was conducted comparing contamination rates when using standard gloves or a new modified product equipped with an additional flap (doffing aid) for easier removal. Gloves were removed after bathing gloved hands in an artificial fluorescent lotion. The number of contamination spots was then visually examined using ultraviolet light. There were 317 individuals who participated in this study: 146 participants (104 nurses and 42 physicians) used standard gloves, whereas 171 participants (118 nurses and 53 physicians) used the modified product. Use of the modified gloves instead of the standard product (15.8% vs 73.3%, respectively; P < .001) and being a physician rather than a nurse (29.5% vs 47.7%, respectively; P = .003) were the only independent risk factors for reduction of contamination. This study shows that the modified product could, at least in vitro, significantly reduce the rate of hand and wrist contamination during removal compared with standard gloves. By this, it may significantly improve the overall quality of patient care when used on the wards directly at the patient's site.


Association between periodontitis and temporomandibular joint disorders.
Periodontitis (PD) may affect temporomandibular joint disorders (TMD) and TMD may influence PD in previous observational studies. Nevertheless, these studies were prone to confounders and reverse causation, leading to incorrect conclusions about causality and direction of association. This research investigates the associations between PD and TMD employing bidirectional two-sample Mendelian randomization (MR) analysis. Single-nucleotide polymorphisms (SNPs) related to PD (p < 5 × 10-6) were selected from a genome-wide association study (GWAS) from the Gene-Lifestyle Interaction in the Dental Endpoints (GLIDE) consortium, and related these to SNPs from FinnGen and UK Biobank (UKB) consortia, and vice versa. We implemented the standard inverse variance weighted (IVW), weighted median (WM), MR-Egger regression, and MR-PRESSO methods to estimate the potential causality between PD and TMD. Sensitive tests were conducted using robust MR methods. Results from FinnGen and UKB were combined using the fixed model. PD did not appear to causally affect TMD. Additionally, the reverse MR analysis did not reveal a significant causal effect of TMD on PD. The results of other MR methods were similar to those of the IVW method. Sensitivity analyses addressed no potential pleiotropy in MR estimations. Results from the meta-analysis were consistent with the above-mentioned consequences. This research does not support a causal relationship between PD and TMD. PD does not appear to worsen TMD directly, and vice versa.


Rhoj Is a Novel Target for Progression and Invasion of Glioblastoma by Impairing Cytoskeleton Dynamics.
Rho GTPase family members were identified as critical regulators of cell morphology, actin cytoskeleton organization, cell movement, and cell cycle and also contributed to tumor progression, which have been implicated in various types of cancer metastasis and growth. Here, we firstly reported the dysregulation of Rhoj in glioblastoma multiforme (GBM) and aimed to investigate the role and mechanism of Rhoj in GBM. We analyzed the expression of 21 Rho GTPases family members and validated the expression of Rhoj in GBM by immunohistochemistry. We further investigated the role and mechanism of Rhoj in GBM both in vitro and in vivo. We observed that Rhoj is significantly overexpressed in GBM and associated with patients' survival. However, the role and underlying molecular mechanism of Rhoj in GBM are still unclear. We demonstrated that transcription factor c-Jun regulated the expression of Rhoj, and Rhoj interacted with moesin to promote GBM cell proliferation and migration by potentiating the activation of Rac1/PAK pathway and cytoskeletal dynamics. Rhoj may promote migration and invasion of GBM cells by regulating epithelial-mesenchymal transition (EMT)-like process. In conclusion, the Rhoj/Rac1/PAK signaling mediates invasion and progression of GBM and is a potential therapeutic target for GBM treatment. Rhoj may also be a promising biomarker for GBM diagnosis and prognosis.


Reproductive effects of synthetic progestin norgestrel in zebrafish (Danio rerio).
The aim of this study was to assess the adverse effects of synthetic progestin norgestrel (NGT) on the reproduction of zebrafish by measuring the egg production, histology and transcriptional expression profiles along the hypothalamic-pituitary-gonadal (HPG) axis in adult zebrafish. After a pre-exposure period of 7 days, adult zebrafish were exposed to 6, 29 and 69 ng L-1 NGT for 21 days. The results showed that exposure to 69 ng L-1 NGT led to a significant up-regulation of follicle stimulating hormone, beta polypeptide (fshb), luteinizing hormone, beta polypeptide (lhb), progesterone receptor (pgr), estrogen receptor 1 (esr1) and androgen receptor (ar) genes in the brains, as well as significant up-regulation of hydroxysteroid 20-beta dehydrogenase (hsd20b) and hydroxysteroid 11-beta dehydrogenase 2 (hsd11b2) genes and down-regulation of 11-beta-hydroxylase (cyp11b) gene in the ovaries of females. In the testes of males, an overall down-regulation of steroidogenic acute regulatory protein (star), cytochrome P450-mediated side-chain cleavage enzyme (cyp11a1), cyp11b, hsd20b, hydroxysteroid 17-beta dehydrogenase type 3 (hsd17b3), hsd11b2 and ar genes were observed following exposure to different treatments of NGT. These transcriptional alterations imply that NGT could exhibit the potent progestogenic and androgenic activities in zebrafish. Egg production as well as histology in the ovaries and testes was not affected by NGT. Taken together, the overall results demonstrated that NGT could significantly affect transcriptional expression levels of genes related to HPG axis in zebrafish, and whether that change translates to additional physiological effects is needed further research.


Altering the inhibitory kinetics and molecular conformation of maltase by Tangzhiqing (TZQ), a natural α-glucosidase inhibitor.
Tangzhiqing (TZQ), as a potential α-glycosidase inhibitor, possesses postprandial hypoglycaemic effects on maltose in humans. The aim of this study was to investigate the mechanisms by which TZQ attenuates postprandial glucose by interrupting the activity of maltase, including inhibitory kinetics and circular dichroism studies. In this study, we determined the inhibitory effect of TZQ on maltase by kinetic analysis to determine the IC50 value and enzyme velocity studies and line weaver-burk plot generation to determine inhibition type. Acarbose was chosen as a standard control drug. After the interaction with TZQ and maltase, secondary structure analysis was conducted with a circular dichroism method. TZQ showed notable inhibition activity on maltase in a reversible and competitive manner with an IC50 value of 1.67 ± 0.09 μg/ml, which was weaker than that of acarbose (IC50 = 0.29 ± 0.01 μg/ml). The circular dichroism spectrum demonstrated that the binding of TZQ to maltase changed the conformation of maltase and varied with the concentration of TZQ in terms of the disappearance of β-sheets and an increase in the α-helix content of the enzyme, similar to acarbose. This work provides useful information for the inhibitory effect of TZQ on maltase. TZQ has the potential to be an α-glycosidase inhibitor for the prevention and treatment of prediabetes or mild diabetes mellitus.


Effects of acute dopamine-agonist treatment in restless legs syndrome on heart rate variability during sleep.
To compare heart rate variability (HRV) changes in patients with restless legs syndrome (RLS) and in healthy subjects, and to evaluate HRV before and after treatment with pramipexole in RLS patients. A prospective, polysomnographic, single-blind, placebo-controlled study was performed in 23 patients with RLS and 10 healthy subjects. Basal spectral analysis of HRV and phasic heart rate (HR) changes during PLMS were compared between the two groups and, within the RLS group, before and after treatment with placebo or pramipexole. No differences were found in the basal sympathovagal balance outside of PLMS between RLS and controls and, in the RLS group, before and after treatment. The amplitude of PLMS-related HR changes was higher in patients than in controls. Treatment with pramipexole decreased the number of PLMS and normalized the increased PLMS-related HR response in RLS subjects. The repetitive abnormal autonomic response to PLMS might play a role in the increased cardiovascular risk proposed for RLS patients. Pramipexole reduced the number of PLMS and the amplitude of the autonomic response to residual PLMS, without effects on the tonic sympathovagal regulation. D3 receptors in the sympathetic pre-ganglionic neurons of the spinal intermediolateral columns might be a target of pramipexole. The normalization of the HR response could be relevant in reducing the risk of cardiovascular diseases and associated autonomic dysfunctions in patients with RLS.


Role of nitric oxide modified DNA in the etiopathogenesis of systemic lupus erythematosus.
The role of the nitric oxide (NO) radical in systemic lupus erythematosus (SLE) pathogenesis has been investigated in the present study. The binding characteristics of SLE autoantibodies with native calf thymus DNA, native and NO-modified plasmid DNA were assessed. Binding characteristics and specificity of antibodies were analysed by direct binding and inhibition ELISA, gel retardation assay and quantitative precipitin titration. The data shows preferential binding of SLE autoantibodies to NO-modified plasmid DNA (NO-DNA) in comparison with native plasmid DNA. Inhibition ELISA reiterates the direct binding results. Gel retardation assay further substantiated the enhanced recognition of NO-DNA by anti-DNA autoantibodies. The binding affinity of modified and native plasmid DNA with one of the SLE IgGs was calculated, using the Langmuir plot. The apparent association constant for NO-plasmid DNA was found to be highest, followed by native calf thymus DNA and native plasmid DNA. The results suggest that the NO radical modification of plasmid DNA causes perturbations, resulting in the generation of neo-epitopes, and making it a potential immunogen. The DNA modified with the NO radical may be one of the factors for the induction of circulating SLE anti-DNA autoantibodies.


Hemicholinium-3 selectively alters the rhythmically bursting activity of septo-hippocampal neurons in the rat.
The medial septal area contains neurons which project to the hippocampal formation. A sizeable proportion of these septo-hippocampal neurons (SHNs) are cholinergic. About 40% of them also display a characteristic discharge pattern in rhythmic bursts. We hypothesized that SHNs with a rhythmically bursting activity (RBA) are the cholinergic ones. To test this hypothesis we studied the effects of acetylcholine synthesis blockade by hemicholinium-3 (HC-3) on the properties of the SHNs. HC-3 (16, 32 or 64 micrograms total dose) or saline were injected in the lateral ventricles of adult male Sprague-Dawley rats anesthetized with urethane. Extracellular recordings from SHNs in the medial septal area were obtained within hours after HC-3 injections (n = 24 animals). SHNs were identified by their antidromic response following electrical stimulation of the fimbria-fornix. The pharmacological properties of SHNs were studied in some animals using microiontophoretic applications from multibarreled electrodes filled with various neurotransmitters. The hippocampal rhythmic slow activity (RSA or theta) was abolished even after the lowest dose of HC-3 tested (16 micrograms). No significant change in SHNs conduction velocity or spontaneous activity was observed at any dose of HC-3. The percentage of SHNs with RBA was unchanged. In contrast the mean frequency of the RBA was decreased by HC-3 in a dose-dependent fashion. The mean frequency was lowest within the first 3 h after injection. Although the mean spontaneous activity was unchanged SHNs tended to have more spikes per burst. The effects of various neurotransmitters on SHNs were qualitatively unchanged after HC-3 injection. These results suggest that acetylcholine synthesis blockade by HC-3 leads not only to the disappearance of the hippocampal RSA in urethane-anesthetized animals, but also to a decrease in the frequency of the rhythmically bursting activity of the SHNs. Since the 4-Hz hippocampal theta is atropine-sensitive, the results provide indirect evidence that the SHNs with rhythmically bursting activity are the cholinergic SHNs.


[Current approaches in multiple myeloma and other cancer-related bone diseases].
Multiple myeloma (MM) ranges second of all hematological malignancies and occurs most commonly in elderly patients. Almost all MM patients develop bone lesions in the course of their disease or have evidence of bone loss at initial diagnosis. Whole-body conventional radiography remains the gold standard in the diagnostic evaluation, albeit computed tomography (CT) and magnetic resonance imaging (MRI) are increasingly used as complementary techniques in the more sensitive detection of osteolytic processes. Bisphosphonates like zoledronate or pamidronate represent the cornerstone therapeutics in osteolytic disease, and are effective supportives to potent anti-myeloma therapies, including novel agents such as the proteasome inhibitor bortezomib or immunomodulatory drugs (IMIDs, e. g. thalidomide or lenalidomide). Several studies are ongoing to investigate the effects of alternative bone-seeking agents and their therapeutic potential for the management of myeloma bone disease, such as denosumab (RANKL-neutralizing antibody), anti-sclerostin (monoclonal antibody, generated against sclerostin) or sotatercept (potent activin-A inhibitor). This review summarizes the most prominent data on myeloma bone disease pathogenesis, the role of imaging techniques as well as therapy and prevention of lytic complications in myeloma which may similarly or equally be true for other bone metastases-inducing solid tumors.


Associations Between Characteristics of Individuals With Fontan Circulation With Blood and Urine Biomarkers of Kidney Injury and Dysfunction.
Background Fontan circulation is associated with kidney injury and dysfunction, often unappreciated until Fontan circulatory failure. We hypothesized that cystatin C-estimated glomerular filtration rate (eGFR) would identify chronic kidney disease more frequently and that urine kidney injury biomarkers would be higher with declining Fontan physiological features. Methods and Results We enrolled 100 ambulatory individuals. Blood and urinary laboratory measurements were compared with demographics and clinically obtained data. Different eGFR equations were used for individuals aged ≥19 years and <19 years. Chronic kidney disease was defined as eGFR <90 mL/min per 1.73 m2. Median (25th-75th percentile) age was 19 (14-26) years, and 43% were female patients. Cystatin C eGFR detected chronic kidney disease (37%) in more patients than creatinine eGFR (11%). Cystatin C eGFR was positively associated, and skeletal muscle mass was negatively associated, with creatinine eGFR in both univariate (cystatin C eGFR β=0.44±0.12, P=0.0006; skeletal muscle mass β=-0.72±0.32, P=0.03) and multivariable analysis (cystatin C eGFR β=0.43±0.12, P=0.0005; skeletal muscle mass β=-0.69±0.29, P=0.02). Urine neutrophil gelatinase-associated lipocalin concentration correlated with Fontan pressure (r=0.28; P=0.04), ventricular end-diastolic pressure (r=0.28; P=0.04), and body fat mass (r=0.26; P=0.03). Conclusions Cystatin C eGFR identified more kidney dysfunction, likely attributable to creatinine eGFR being confounded by skeletal muscle mass. Elevated urine neutrophil gelatinase-associated lipocalin was associated with worse Fontan hemodynamics and higher percentage body fat, suggesting that higher venous pressure and higher adiposity are associated with ongoing kidney injury.


The affection on the tissue lesions of difference frequency in dual-frequency high-intensity focused ultrasound (HIFU).
The lesions induced by dual-frequency high-intensity focused ultrasound (HIFU) in freshly excised porcine livers were investigated and compared with the lesions induced by conventional single-frequency HIFU. The results have shown that using different exposure time resulted in lesions of different sizes in both dual-frequency and single-frequency HIFU modes at the same intensity level (ISAL = 808 W cm(-2)), but the dimensions of lesions in dual-frequency mode were obviously larger than those in single-frequency mode. A much possible explanation was suggested for the results and the possibilities of applications in the future were also discussed in brief.


Experimental human exposure to carbon disulfide. II. Urinary excretion of 2-thiothiazolidine-4-carboxylic acid (TTCA) during and after exposure.
Six human volunteers were exposed to 10 and 20 ppm carbon disulfide at rest and to 3 and 10 ppm carbon disulfide under a 50 W level of physical exercise during four consecutive periods of 50 min. At the start of the experiments, at the end of the exposure periods and during the post-exposure period, urine was sampled and the concentration of 2-thiothiazolidine-4-carboxylic acid (TTCA) was determined. It was established that only a small percentage, ranging from 0.7 to 2.2% of the absorbed carbon sulfide was transformed into TTCA. The excretion rate of TTCA (mumol TTCA h-1) was found to be the best parameter in evaluating the respiratory uptake of carbon disulfide over a range of 37.9 to 163.3 mg CS2 compared to the urinary concentration of TTCA (mole TTCA ml-1) or the creatinine corrected concentration of TTCA (mmol TTCA mol-1 creatinine). The total amount of TTCA (mumol TTCA) excreted proved to be independent of the urinary flow (ml h-1), the estimates of the individual fatty tissue content and the urinary pH. No correlation was found between the respiratory uptake of carbon disulfide (mg CS2) and the excretion rate of TTCA within each exposure condition of 3, 10 or 20 ppm carbon disulfide, respectively.


Macroglossia secondary to systemic amyloidosis: case report and literature review.
Amyloidosis is characterized by an abnormal extracellular deposition of amyloid in different tissues and organs, where it usually causes some type of dysfunction. Its cause is unknown. The two main forms of amyloidosis are systemic and localized; the latter is rare. No satisfactory treatment for systemic amyloidosis has been discovered, and mean survival is poor, ranging from 5 to 15 months depending on the presence or absence of multiple myeloma. We report a case of primary systemic amyloidosis in a 71-year-old man. The diagnosis of amyloidosis was established by tongue biopsy, and its systemic nature was identified by analysis of aspirated abdominal fat. At the 1-year follow-up, the patient's clinical condition had not changed, and he was thereafter lost to follow-up.


Marginal discrepancy of 3-unit Co-Cr metal copings fabricated with additive and subtractive manners: A comparative study.
Poor marginal adaptation may give rise to a series of biological complications. Despite its importance, comparative studies on marginal adaptation of metal-ceramic fixed restorations fabricated with newer methods are limited. Therefore, it was aimed to assess the marginal accuracy of copings fabricated with subtractive and additive manners used in contemporary dentistry. For a typodont model, 60 three-unit cobalt-chrome metal copings were fabricated by selective laser sintering (SLS), hard alloy milling (HAM), and soft alloy milling (SAM) in comparison to casting (C). Marginal discrepancy assessment was performed by using replication technique. Mesiodistal and buccopalatal cross-sections of silicone replicas were investigated under a stereomicroscope at × 80 magnification. A total of 960 measurements were subsequently made by means of corresponding image-review software on captured images after calibration of the software to μm scale. Obtained data were subjected to nonparametric Kruskal Wallis and Tamhane post-hoc tests (α =0.05). Marginal adaptation of C group was significantly inferior to test groups in both canine- and premolar-teeth measurements (P < 0.05). Marginal fit was found to be tooth dependent (P < 0.001). HAM, SAM, and SLS groups exhibited analogous marginal discrepancy values on canine-tooth measurements. Differences among these groups were not statistically significant (P > 0.05). SAM and SLS groups demonstrated better marginal adaptation than others on premolar-tooth measurements. Also, no statistically significant difference was detected between SAM and SLS groups (P > 0.05). SAM group demonstrated superior marginal accuracy. All groups had clinically acceptable marginal discrepancy values (<120 μm), except cast group.


Bilateral testicular Leydig cell hyperplasia presented incidentally: A case report.
Leydig cell hyperplasia or tumor represents less than 3% of all testicular tumors. It can be defined as an increase in the size and number of Leydig cell within the testicles. These cells are responsible for the production of testosterone in human males. Our patient is a forty-eight-year-old male presented with erectile dysfunction and decreased libido for the past six months. Ultrasound of the scrotum show bilateral hypoechoic testicular masses larger on the left size thus left orchidectomy was performed. Histopathology confirmed our diagnosis. Leydig cell hyperplasia (LCH) is a rare and mostly benign entity that affects both children and adults. In adults, it might be associated with variety of condition including Klinefelter's syndrome, exogenous human chorionic gonadotropin (hCG) therapy, and many others but it mostly occurs idiopathically. Scrotal ultrasound and tumor markers can be used to diagnose most of the patients with LCH. LCH should be differentiated from Leydig cell tumor to avoid unnecessary and sometimes harmful intervention in the future.


Capillary rarefaction characteristic of the skeletal muscle of hypertensive patients.
There is evidence that the rarefaction of the capillary bed is typical for the skeletal muscle of spontaneously hypertensive rats. We were therefore interested to learn whether there is also a rarefaction in skeletal muscle of human hypertensives. The number of capillaries was morphometrically analysed and counted in the quadriceps and the pectoralis major muscles of human normotensives (n = 12) and hypertensives (n = 15). The clinical diagnosis and certain pathological criteria, such as blood pressure (with or without antihypertensive therapy), heart weight, left ventricular wall thickness, the state of kidney arterioles and brain, and heart vessels, were used to classify the patients into two groups. The dissected tissue samples were prepared according to the GMA method and the capillary numbers per area were counted using light microscopy (250 x). The quadriceps muscle had a capillary density (per 2.5 mm2) of 442 +/- 51 in normotensives and 277 +/- 41 in hypertensive patients; in the pectoralis major muscle we counted 477 +/- 30 in controls and 232 +/- 28 in hypertensives. The rarefaction in the quadriceps muscle ranged by about 37%, in the pectoralis major muscle by about 51%. It is suggested that the reduction of the capillary surface area caused by the capillary rarefaction reduces the transcapillary fluid exchange and in that way prevents an overperfusion of the terminal vascular bed.


Neurokinin-induced changes in pial artery diameter in the anaesthetized guinea-pig.
1. The effects of selective neurokinin agents on pial artery diameter, measured with an on-line image analyser, have been studied in anaesthetized guinea-pigs in order to characterize the neurokinin receptors present on pial arteries. 2. Perivascular injection of either substance P (0.01-1 microM) or the selective NK1 receptor agonists, substance P methyl ester (SPOMe, 0.01-1 microM) and GR73632 (0.1 microM), increased pial artery diameter. 3. In contrast, the selective NK2 receptor agonist, GR64349 (1 microM), produced a small vasoconstriction while the NK3 receptor-selective agonist, senktide (1 microM) was inactive. 4. Co-administration of GR82334 (1 microM), a selective NK1 receptor antagonist, inhibited the vasodilatation produced by SPOMe (0.1 microM) but not that caused by calcitonin gene-related peptide (CGRP, 0.01 microM). 5. The results are consistent with an involvement of NK1 receptors in the neurokinin-induced increase in guinea-pig pial artery diameter.


Organochlorine and butyltin residues in walleye pollock (Theragra chalcogramma) from Bering Sea, Gulf of Alaska and Japan Sea.
Persistent organochlorine (OC) and toxic butyltin compounds (BTs) were determined in walleye pollock (Theragra chalcogramma) collected from Gulf of Alaska, Bering Sea and Japan Sea, during 1991 and 1992. Polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDTs) and its metabolites were the most abundant compounds ranging up to 3200 and 2500 ng/g on lipid weight, respectively, followed by chlordane compounds (CHLs), hexachlorocyclohexane isomers (HCHs) and hexachlorobenzene (HCB) in the liver of walleye pollock. Concentrations of HCHs and HCB in walleye pollock from these remote areas were higher than those in fishes from the western North Pacific and Japanese coastal waters, indicating atmospheric transport of these compounds to higher latitude regions such as Bering Sea and Gulf of Alaska and/or local input around northern Japan Sea. The concentrations of other OCs were generally comparable to those in fishes from North Pacific Ocean and Japanese waters but significantly lower than in cod-like fishes from North Atlantic and European countries. Among sampling locations, walleye pollock from Japan Sea showed higher concentrations of DDTs and HCHs compared to fishes from Bering Sea and Gulf of Alaska, suggesting greater input of these compounds around Japan Sea. Slower declining trend of DDTs and CHLs and an increasing pattern of PCBs concentrations were found in walleye pollock from Bering Sea during 1982-1992. This may imply a continuous input of these compounds by long-range transport and/or long-term persistency in these cold regions. Compared to the fishes from Japan Sea, walleye pollock from Bering Sea and Gulf of Alaska showed higher proportions of alpha-HCH and p,p'-DDE in the composition of HCH isomers and DDT compounds, respectively. This suggests selective transportability of these compounds during long-range transport to higher latitude remote areas. Concentrations of tributyltin (TBT) in the muscle of walleye pollock ranged from 1.1 to 5.5 ng/g on wet weight. Concentrations of TBT in deep-sea walleye pollock from Gulf of Alaska and Bering Sea were lower than those in Japan Sea.


Overexpression of cyclo-oxygenase-2 is an independent predictor of unfavourable outcome in node-negative breast cancer, but is not associated with protein kinase B (Akt) and mitogen-activated protein kinase (ERK1/2, p38) activation or with Her-2/neu signalling pathways.
The production of prostaglandins is regulated by cyclo-oxygenases (COXs), which also have a role in tumour development and progression in various malignancies, including breast cancer. The mechanisms by which COX-2 contributes to unfavourable prognosis are still poorly understood. The association between expression of COX-2 and possible linked signalling pathways-namely, Akt, extracellular regulated kinases (ERK1/2), the stress-activated kinase p38 or Her-2/neu-is assessed in a series of 113 node-negative breast cancers. COX-2 was identified as an independent prognostic factor (p = 0.034) in node-negative breast cancer by survival analysis. The lack of a relationship between COX-2 expression and activated Akt, Erk1/2, p38 and Her-2/neu was indicated by statistical analysis. The prognostic effect of COX-2 expression on lymph node-negative breast cancer is confirmed-COX-2 is probably not regulated by HER-2, Akt, Erk1/2 or p38. Further studies are necessary for the elucidation of the signalling pathways responsible for the modification of COX-2 expression and the increased aggressiveness of breast cancers overexpressing COX-2.


A voltage trade study for the design of capillary electrophoresis instruments for spaceflight.
Capillary electrophoresis (CE) systems have undergone extensive development for spaceflight applications. A flight-compatible high voltage power supply and the necessary voltage isolation for other energized components can be large contributors to both the volume and mass of a CE system, especially if typical high voltage levels of 25-30 kV are used. Here, we took advantage of our custom CE hardware to perform a trade study for simultaneous optimization of capillary length, high voltage level, and separation time, without sacrificing method performance. A capillary electrophoresis with capacitively coupled contactless conductivity detection (CE-C4 D) method recently developed by our group to target inorganic cations and amino acids relevant to astrobiology was used as a test case. The results indicate that a 50 cm long capillary with 15 kV applied voltage (half of that used in the original method) can be used to achieve measurement goals while minimizing instrument size.


Clinical application of 62Zn/62Cu positron generator: perfusion and plasma pool images in normal subjects.
We have developed a new 62Zn/62Cu positron generator, and applied it for PET imaging of perfusion and plasma volume in 5 normal subjects. The generator makes it possible by a simple procedure to obtain 62Cu eluate and labeling compounds sufficiently every 40-60 minutes. 62Cu labeled pyruvaldehyde bis(N4-methylthiosemicarbazone) copper II (62Cu-PTSM) was employed for cerebral and myocardial perfusion imaging and 62Cu labeled human serum albumin-dithiosemicarbazone (62Cu-HSA-DTS) was used for plasma pool imaging. The images of cerebral blood flow, cerebral plasma volume and myocardial perfusion were excellent. In addition, the analysis of tissue activity and blood activity demonstrated the microspheric character of 62Cu-PTSM. Correction of arterial activity with the standard disappearance curve of 62Cu-PTSM suggested the possibility of quantifying blood flow. The results of this study indicate the capability of the 62Zn/62Cu generator for wide clinical use without an in-house cyclotron.


Everolimus-induced somatostatin receptor overexpression in a rectal neuroendocrine tumor patient may promote somatostatin receptor-guided radionuclide therapy (peptide receptor radiotherapy) as an additional treatment option.
We present a case of Grade II, well-differentiated rectal neuroendocrine tumor in a 39-year-old patient. Following different sequential treatment modalities, the disease progressed both on metabolic (18F-fluorodeoxyglucose positron emission tomography-computed tomography [18F-FDG PET/CT]) and somatostatin receptor (SSTR)-imaging (68Ga-DOTA-Tyr3-octreotate [68Ga-DOTATATE] PET/CT), and the patient received three cycles of peptide receptor radiotherapy (PRRT). Two years later, upon new progression due to the appearance of metabolically active, 68Ga-DOTATATE PET/CT-negative liver lesions, targeted treatment with everolimus was introduced. Further morphologic and metabolic progression occurred 4 months after everolimus initiation, however, this time liver lesions demonstrated increased SSTR-expression on68Ga-DOTATATE PET/CT. Thus, the patient became eligible for a second PRRT course.


Immunomodulatory effects of testosterone evaluated in all-trans retinoic acid differentiated HL-60 cells, granulocytes, and monocytes.
The sex hormones are known to affect innate immunity in humans. In this study we evaluated the immunomodulatory effects of testosterone in a model system comprising of all-trans retinoic acid differentiated HL-60 cells, and confirmed the results in human granulocytes and monocytes. Results showed that testosterone at pharmacological doses reduced the production of interleukin-8 and reactive oxygen species from differentiated HL-60 cells in a concentration dependent manner without affecting phagocytosis. The cells were stimulated with zymosan, lipopolysaccharide, or Bacillus subtilis. At the highest concentration of testosterone (120 μM), interleukin-8 secretion was reduced 42-80%, and production of reactive oxygen species was reduced 32-46%. Flutamide, an antagonist of the classical intracellular androgen receptor, was unable to antagonize the immunosuppressive effect of testosterone. We further demonstrated that the suppressive effect of testosterone has a short onset time. Our results suggest that testosterone affects the fast operating membrane bound androgen receptor or a rapid acting enzyme system. Testosterone, at pharmacological doses, was also shown to suppress generation of reactive oxygen species and interleukin-8 in human granulocytes and monocytes, respectively, to a similar extent as observed in differentiated HL-60 cells.


Ablation of Ca2+ channel beta3 subunit leads to enhanced N-methyl-D-aspartate receptor-dependent long term potentiation and improved long term memory.
The beta subunits of voltage-dependent Ca(2+) channels (VDCCs) have marked effects on the properties of the pore-forming alpha(1) subunits of VDCCs, including surface expression of channel complexes and modification of voltage-dependent kinetics. Among the four different beta subunits, the beta(3) subunit (Ca(v)beta3) is abundantly expressed in the hippocampus. However, the role of Ca(v)beta3 in hippocampal physiology and function in vivo has never been examined. Here, we investigated Ca(v)beta3-deficient mice for hippocampus-dependent learning and memory and synaptic plasticity at hippocampal CA3-CA1 synapses. Interestingly, the mutant mice exhibited enhanced performance in several hippocampus-dependent learning and memory tasks. However, electrophysiological studies revealed no alteration in the Ca(2+) current density, the frequency and amplitude of miniature excitatory postsynaptic currents, and the basal synaptic transmission in the mutant hippocampus. On the other hand, however, N-methyl-d-aspartate receptor (NMDAR)-mediated synaptic currents and NMDAR-dependent long term potentiation were significantly increased in the mutant. Protein blot analysis showed a slight increase in the level of NMDAR-2B in the mutant hippocampus. Our results suggest a possibility that, unrelated to VDCCs regulation, Ca(v)beta3 negatively regulates the NMDAR activity in the hippocampus and thus activity-dependent synaptic plasticity and cognitive behaviors in the mouse.


Reoperation after early and late failure of mitral valve repair for degenerative disease.
To determine mechanisms of early and late failure after mitral valve repair for degenerative disease, identify factors associated with re-repair, and evaluate durability of re-repair. From January 2008 to July 2020, 330 reoperations were performed for recurrent mitral valve dysfunction after initial valve repair for degenerative disease. Mechanisms of repair failure were determined by review of preoperative imaging and operative reports. Multivariable analysis was performed to identify factors associated with likelihood of re-repair or replacement. Durability of re-repair was assessed using longitudinal analysis of postoperative echo data. Eighty-five of 330 (26%) reoperations were performed for early repair failure within 1 year and 245/330 (74%) for late failure thereafter. Suture/annuloplasty dehiscence, systolic anterior motion, hemolysis, and ventricular remodeling were more common mechanisms of early failure and disease progression and fibrosis late failure. Forty percent (34/85) of early failures were re-repaired versus 24% (59/245) of late failures. Re-repair was more common than replacement in recent years and was associated with earlier reoperation (median 1.5 vs 3.9 years; P = .0001). No in-hospital deaths occurred after re-repair; 2 patients (0.8%) died after valve replacement. Freedom from severe mitral regurgitation after re-repair was 93% at 7 years. Mitral valve re-repair can be performed with low rates of mortality and morbidity for early and late valve dysfunction. Mechanisms of early repair failure differ from those of late failure and are generally more amenable to re-repair. In selected patients who present after failed repair, we prefer mitral re-repair to valve replacement whenever technically feasible.


Efficacy and Safety of Corticosteroid Implants in Non-infectious Uveitis: A Systematic Review with Network Meta-analysis.
To compare different existent implants for noninfectious uveitis. We performed a systematic review of randomized clinical trials (RCTs), searching in five electronic databases and screening reference lists of included studies and relevant reviews. We performed network meta-analysis to compare 0.18 mg fluocinolone acetonide implants, 0.70 mg and 0.35 mg dexamethasone implants, and sham procedures. Eight RCTs were included in this systematic review, of which three articles from two studies (n = 358 patients) were included in network meta-analysis. The 0.70 mg dexamethasone implant (at 1.5 months) was associated with better results than the 0.18 mg fluocinolone acetonide implant (at 1 month) at improving vitreous haze grading (RR = 2.96; 95%CI = 1.23-7.07), and with less frequently development of cataracts at 12 (RR = 0.36; 95%CI = 0.17; 0.79) and 36 months (RR = 0.37; 95%CI = 0.20; 0.71). In the short term, the 0.70 mg dexamethasone implant is superior at improving vitreous haze, and shows lower risk of cataracts. However, additional RCTs with standardized outcomes are needed.


Brassica rapa Polysaccharides Ameliorate CCl4 -Induced Acute Liver Injury in Mice through Inhibiting Inflammatory Apoptotic Response and Oxidative Stress.
Brassica rapa L., also called NIUMA, is used empirically in Tibetan medicine for its antioxidant, anti-inflammatory and antiradiation activities. This study explored the hepatoprotective effects of B. rapa polysaccharides (BRPs) on acute liver injury induced by carbon tetrachloride (CCl4 ) in mice and the underlying mechanisms. Mice were treated with CCl4 after the oral administration of BRPs (55, 110 and 220 mg/kg) or bifendate (100 mg/kg) for 7 days. Blood and liver samples of mice were collected for analysis after 24 h. The ALP, ALT and AST levels and the biological activities of SOD, MDA and GSH-Px were measured. Histopathological changes in the liver were determined through hematoxylin and eosin staining. Moreover, TNF-α, IL-1β and IL-6 expression levels were detected by commercial reagent kits. Finally, Western blot analysis was used to check the relative expression levels of caspase-3, p-JAK2 and p-STAT3. The BRP pre-treatment significantly decreased the enzymatic activities of ALT, ALP and AST in the serum, markedly increased the activities of SOD and GSH-Px in the liver and reduced the MDA concentration in the liver. BRPs alleviated hepatocyte injury and markedly inhibited the expression of TNF-α, IL-1β and IL-6, also downregulating the CCl4 -induced hepatic tissue expression of caspase-3. Furthermore, BRPs inhibited the JAK2/STAT3 signaling pathway in a dose-dependent manner in the liver. This study demonstrated that BRPs exert hepatoprotective effect against the CCl4 -induced liver injury via modulating the apoptotic and inflammatory responses and downregulating the JAK2/STAT3 signaling pathway. Therefore, B. rapa could be considered a hepatoprotective medicine.


One-step sandwich enzyme immunoassay for human laminin using monoclonal antibodies.
A one-step sandwich enzyme immunoassay (one step sandwich EIA) for human serum immunoreactive laminin was set up with a pair of monoclonal antibodies prepared against human placental laminin P1 fragment. The assay was characterized by carrying out two immunoreactions simultaneously, laminin P1 fragment reacting with both a monoclonal antibody as a solid phase and a horseradish peroxidase-labeled monoclonal antibody (Fab') against human laminin P1 fragment as conjugate. Sensitivity of the immunoassay was 0.01 ng/well (0.5 microgram/l), and linearity was obtained between 0.01-20 ng/well (0.5-1,000 micrograms/l). The levels of laminin in sera from normal individuals and patients with liver cirrhosis, hepatocellular carcinoma and primary biliary cirrhosis were 103 +/- 15 micrograms/l, 228 +/- 70 micrograms/l, 341 +/- 163 micrograms/l and 232 +/- 93 micrograms/l, respectively. Protein immunoblotting showed that the serum immunoreactive laminin measured by the assay was a fragment with rel mol mass of 200 kDa.


[Head and neck adaptive radiotherapy].
Onboard volumetric imaging systems can provide accurate data of the patient's anatomy during a course of head and neck radiotherapy making it possible to assess the actual delivered dose and to evaluate the dosimetric impact of complex daily positioning variations and gradual anatomic changes such as geometric variations of tumors and normal tissues or shrinkage of external contours. Adaptive radiotherapy is defined as the correction of a patient's treatment planning to adapt for individual variations observed during treatment. Strategies are developed to selectively identify patients that require replanning because of an intolerable dosimetric drift. Automated tools are designed to limit time consumption. Deformable image registration algorithms are the cornerstones of these strategies, but a better understanding of their limits of validity is required before adaptive radiotherapy can be safely introduced to daily practice. Moreover, strict evaluation of the clinical benefits is yet to be proven.


Sudden infant death syndrome and the modulation of neuropeptides released during shock.
It is generally accepted that sudden infant death syndrome (SIDS) victims fail to survive relatively minor stress in infancy. My hypothesis is that failure to orchestrate the endocrine response in stress leads to excessive release of neutral-endopeptidase-sensitive peptide substrates that enhance lethality. The 'quick zinc' response Reid recorded in livestock with circulatory shock is described. It is concluded that the failure to mount an endocrine response leads to SIDS.


Symptomatic venous thromboembolism in Chinese patients after gynecologic surgery: incidence and disease pattern.
Venous thromboembolism is a common cause of postoperative morbidity and mortality in Caucasian populations, but it is widely believed that this complication is rare in Chinese. We conducted a retrospective study from January 1998 to December 2000. Women with thromboembolic diseases after gynecologic surgery were identified and their medical records were reviewed. During the study period, thirty-one women were diagnosed as having thromboembolic disease after gynecologic surgery. Over the study period, the total number of operations was 6077, giving an incidence of 5.10/1000 operations. There were two cases of pulmonary embolism and the others had deep vein thrombosis of which 90% were limited to calf veins only. The incidences were significantly higher in 1999 and 2000 (7.59 and 6.85/1000 operations, respectively) than that in 1998 (1.7/1000 operations) (P = 0.015), after a case of maternal death due to pulmonary embolism in early 1999. Most cases of thromboembolism were diagnosed after major surgery for malignancy (n = 14) or benign conditions (n = 12). In the remaining cases, three had evacuation of the uterus for retained products of conception and two had laparoscopy for suspected ectopic pregnancies. Thromboembolic disease is not uncommon among Chinese women after gynecologic surgery. The incidence is similar to that of the Caucasian population, although the sites of vascular occlusion were different. The long belief that thromboembolism is rare among Chinese is at least partly due to under-diagnosis.


Near-miss analysis in a large hospital in southern Brazil: A 5-year retrospective study.
Near-miss analysis is an effective method for preventing serious adverse events, including never events such as wrong-site surgery. To analyze all near-miss incidents reported in a large general hospital in southern Brazil between January 2013 and August 2017. We performed a descriptive retrospective study of near-miss incidents recorded in the hospital's electronic reporting system in a large non-profit hospital (497 beds). The results are expressed as absolute (n) and relative frequencies (%). Pearson's chi-square test, Fisher's exact test (Monte Carlo simulation) and linear regression were used. A total of 12,939 near-miss incidents were recorded during the study period, with linear growth in the number of reports. Near-miss incidents were most frequent for medication, followed by processes unspecified in the International Classification for Patient Safety framework, followed by information control (patient chart and fluid balance data), followed by venous/vascular puncture. The highest prevalence of reports was observed in inpatient wards, in adult, pediatric, and neonatal intensive care units, and in the surgical center/post-anesthesia care unit. Pharmacists and nursing personnel recorded most of the reports during the day shift. The most frequent categories of near-miss incidents were medication processes, other institutional protocols, information control issues, and venous/vascular puncture. The significant number of reported near-miss incidents reflects good adherence to the reporting system.


Peripheral nerve injury-induced alterations in VTA neuron firing properties.
The ventral tegmental area (VTA) is one of the main brain regions harboring dopaminergic (DA) neurons, and plays important roles in reinforcement and motivation. Recent studies have indicated that DA neurons not only respond to rewarding stimuli, but also to noxious stimuli. Furthermore, VTA DA neurons undergo plasticity during chronic pain. Lateral and medial VTA neurons project to different brain areas, and have been characterized via their distinct electrophysiological properties. In this study, we characterized electrophysiological properties of lateral and medial VTA DA neurons using DAT-cre reporter mice, and examined their plasticity during neuropathic pain states. We observed various DA subpopulations in both the lateral and medial VTA, as defined by action potential firing patterns, independently of synaptic inputs. Our results demonstrated that lateral and medial VTA DA neurons undergo differential plasticity after peripheral nerve injury that leads to neuropathic pain. However, these changes only reside in specific DA subpopulations. This study suggests that lateral and medial VTA DA neurons are differentially affected during neuropathic pain conditions, and emphasizes the importance of subpopulation specificity when targeting VTA DA neurons for treatment of neuropathic pain.


Protection of Escherichia coli K12 structural integrity by Ca2+ and Mg2+ in bactericidal and bacteriolytic tests.
Escherichia coli K12 suspended in different media showed a loss of phospholipids. Mg2+ and Ca2+ 0.01 M prevented phospholipid loss and stabilized Escherichia coli K12 for bactericidal and bacteriolytic assays.


Gender differences of pulsed and tissue Doppler indexes of left ventricular diastolic function in healthy subjects.
The purpose of this study is to determine the normative Doppler values and gender differences in left ventricular (LV) diastolic function in healthy subjects at each decade of life. Four hundred sixty-seven male and 455 female healthy subjects, aged 20-90 years, underwent standard echocardiography and tissue Doppler imaging. We divided the subjects into seven groups, based on the decade of their age (i.e., 20s, 30s, 40s, 50s, 60s, 70s, and 80s), and measured standard echocardiographic parameters and Doppler indices. Females younger than 50 years had significantly higher mitral E velocity, and females older than 40 years had significantly higher mitral A velocity, compared with their male counterparts. There was no difference in mitral E to A velocity ratio (E/A) at any age. The tissue Doppler method showed significantly lower early diastole velocity of the mitral annulus (E a) in females-especially females older than 50 years-than in males (10.7 ± 3.7 versus 11.2 ± 3.7 cm/s, p < 0.025). Mean E/E a ratio was higher in females than in males (6.9 ± 2.4 versus 6.0 ± 2.0, p < 0.0001). Our results indicate gender differences in Doppler indexes of LV diastolic function in healthy subjects. In clinical settings, assessment of LV diastolic function using the Doppler method should take into account patient gender.


Differences in Failure-Free Survival After Salvage Radiotherapy Guided by Conventional Imaging Versus 18F-Fluciclovine PET/CT in Postprostatectomy Patients: A Post Hoc Substratification Analysis of the EMPIRE-1 Trial.
The EMPIRE-1 (Emory Molecular Prostate Imaging for Radiotherapy Enhancement 1) trial reported a survival advantage in recurrent prostate cancer salvage radiotherapy (SRT) guided by 18F-fluciclovine PET/CT versus conventional imaging. We performed a post hoc analysis of the EMPIRE-1 cohort stratified by protocol-specified criteria, comparing failure-free survival (FFS) between study arms. Methods: EMPIRE-1 randomized patients to SRT planning via either conventional imaging only (bone scanning plus abdominopelvic CT or MRI) (arm A) or conventional imaging plus 18F-fluciclovine PET/CT (arm B). Randomization was stratified by prostate-specific antigen (PSA) level (<2.0 vs. ≥ 2.0 ng/mL), adverse pathology, and androgen-deprivation therapy (ADT) intent. We subdivided patients in each arm using the randomization stratification criteria and compared FFS between patient subgroups across study arms. Results: Eighty-one and 76 patients received per-protocol SRT in study arms A and B, respectively. The median follow-up was 3.5 y (95% CI, 3.0-4.0). FFS was 63.0% and 51.2% at 36 and 48 mo, respectively, in arm A and 75.5% at both 36 and 48 mo in arm B. Among patients with a PSA of less than 2 ng/mL (mean, 0.42 ± 0.42 ng/mL), significantly higher FFS was seen in arm B than arm A at 36 mo (83.2% [95% CI, 70.0-91.0] vs. 66.5% [95% CI, 51.6-77.8], P < 0.001) and 48 mo (83.2% [95% CI, 70.0-91.0] vs. 56.2% [95% CI, 40.5-69.2], P < 0.001). No significant difference in FFS between study arms in patients with a PSA of at least 2 ng/mL was observed. Among patients with adverse pathology, significantly higher FFS was seen in arm B than arm A at 48 mo (68.9% [95% CI, 52.1-80.8] vs. 42.8% [95% CI, 26.2-58.3], P < 0.001) though not at the 36-mo follow-up. FFS was higher in patients without adverse pathology in arm B versus arm A (90.2% [95% CI, 65.9-97.5] vs. 73.1% [95% CI, 42.9-89.0], P = 0.006) at both 36 and 48 mo. Patients in whom ADT was intended in arm B had higher FFS than those in arm A, with the difference reaching statistical significance at 48 mo (65.2% [95% CI, 40.3-81.7] vs. 29.1 [95% CI, 6.5-57.2], P < 0.001). Patients without ADT intent in arm B had significantly higher FFS than patients in arm A at 36 mo (80.7% [95% CI, 64.9-90.0] vs. 68.0% [95% CI, 51.1-80.2]) and 48 mo (80.7% [95% CI, 64.9-90.0] vs. 58.6% [95% CI, 41.0-72.6]). Conclusion: The survival advantage due to the addition of 18F-fluciclovine PET/CT to SRT planning is maintained regardless of the presence of adverse pathology or ADT intent. Including 18F-fluciclovine PET/CT to SRT leads to survival benefits in patients with a PSA of less than 2 ng/mL but not in patients with a PSA of 2 ng/mL or higher.


Identification of insulin-regulated aminopeptidase (IRAP) in the rat pineal gland and the modulation of melatonin synthesis by angiotensin IV.
A local renin-angiotensin system (RAS) has been postulated in the pineal gland. In addition to angiotensin II (Ang II), other active metabolites have been described. In this study, we aimed to investigate a role for Ang IV in melatonin synthesis and the presence of its proposed (IRAP)/AT4 receptor (insulin-regulated aminopeptidase) in the pineal gland. The effect of Ang IV on melatonin synthesis was investigated in vitro using isolated pinealocytes. IRAP protein expression and activity were evaluated by Western blot and fluorimetry using Leu-4Me-β-naphthylamide as a substrate. Melatonin was analyzed by HPLC, calcium content by confocal microscopy and cAMP by immunoassay. Ang IV significantly augmented the NE-induced melatonin synthesis to a similar degree as that achieved by Ang II. This Ang IV effect in pinealocytes appears to be mediated by an increase in the intracellular calcium content but not by cAMP. The (IRAP)/AT4 expression and activity were identified in the pineal gland, which were significantly higher in membrane fractions than in soluble fractions. Ang IV significantly reduced IRAP activity in the pineal membrane fractions. The main findings of the present study are as follows: (1) Ang IV potentiates NE-stimulated melatonin production in pinealocytes, (2) the (IRAP)/AT4 receptor is present in the rat pineal gland, and (3) Ang IV inhibits IRAP activity and increases pinealocytes [Ca2+]i. We conclude that Ang IV is an important component of RAS and modulates melatonin synthesis in the rat pineal gland.


The Ecological Differentiation of Particle-Attached and Free-Living Bacterial Communities in a Seasonal Flooding Lake-the Poyang Lake.
Particle-attached (PA) and free-living (FL) bacterial communities play essential roles in the biogeochemical cycling of essential nutrients in aquatic environments. However, little is known about the factors that drive the differentiation of bacterial lifestyles, especially in flooding lake systems. Here we assessed the compositional and functional similarities between the FL and PA bacterial fractions in a typical flooding lake-the Poyang Lake (PYL) of China. The results revealed that PA communities had significantly different compositions and functions from FL communities in every hydrological period, and the diversity of both PA and FL communities was affected mainly by the water regime rather than bacterial lifestyles. PA communities were more diverse and enriched with Proteobacteria and Bacteroidetes, while FL communities had more Actinobacteria. There was a higher abundance of photosynthetic and nitrogen-cycling bacterial groups in PA communities, but a higher abundance of members involved in hydrocarbon degradation, aromatic hydrocarbon degradation, and methylotrophy in FL communities. Water properties (e.g., temperature, pH, total phosphorus) significantly regulated the lifestyle variations of PA and FL bacteria in PYL. Collectively, our results have demonstrated a clear ecological differentiation of PA and FL bacterial communities in flooding lakes, suggesting that the connectivity between FL and PA bacterial fractions is water property-related rather than water regime-related.


An amperometric acetylcholine biosensor based on a conducting polymer.
An amperometric acetylcholine biosensor was prepared by the generation of the conducting polymer poly(4-(2,5-di(thiophen-2-yl)-1H-pyrrol-1-yl)benzenamine) (poly(SNS-NH2)) on graphite electrodes. For pesticide detection, the enzymes acetylcholinesterase (AChE) and choline oxidase (ChO) were co-immobilized onto the conducting polymer poly(SNS-NH2) films using covalent binding technique. Electrochemical polymerization was carried out using a three-electrode cell configuration via cyclic voltammetry. Characterization of resulting acetylcholine biosensor was done in terms of optimum pH, enzyme loading, range of linear response and shelf-life. Linear range was 0.12-10mM and shelf-life 4 weeks. Sensitivity was calculated as 2.19μAmM(-1)cm(-2). The designed biosensor was tested for the determination of paraoxon-ethyl in spiked tap water samples. The results were compared with a conventional quantification method using HPLC-DAD. Linear correlation of the quantification results with both methods (R(2)=0.998) was obtained.


Factors contributing to the decline of leprosy in Spain in the second half of the twentieth century.
Leprosy is a chronic infectious disease that is considered to be declining, though it still remains prevalent in many parts of the world. A study was made to explore the health and socioeconomic factors that most influenced the trend of the disease in a typical Mediterranean country. An ecological study was conducted, investigating possible social, economic and health factors related to the evolution of leprosy incidence. The time period considered was 50 years--the second half of the twentieth century in Spain. The variables showing the strongest correlation to evolution of the incidence of the disease were employment, the number of physicians, and the gross domestic product (GDP), with negative coefficients--while tuberculosis showed a positive coefficient. However, the GDP showed the highest coefficient (0.5). The model that best explained the evolution of leprosy over the last 50 years comprised a 6-year lag period between the socioeconomic factors and the incidence of leprosy--explaining 57% of the data obtained. The annual decrease in leprosy incidence was 1.6%. Socioeconomic development, assessed in terms of the GDP, was the most important factor in explaining the evolution of leprosy incidence.


Impact of ISO/IEC 17025 laboratory accreditation in sub-Saharan Africa: a case study.
The number and severity of nonconformities generated during an audit of a medicine testing laboratory indicates its level of quality compliance. Quality standards are established and maintained to ensure the reliability of laboratory test reports. The National Medicines Regulatory Authority (NMRA) Quality Control laboratories assess the quality of medicines used by the populace as part of their regulatory function. Although countries desire to have reliable medicine testing facilities, accrediting a national laboratory to international standards poses financial and technical challenges for many low-income countries. Sharing the benefits of laboratory accreditation could help more countries within sub-Saharan Africa overcome existing challenges to achieve accreditation and robust quality systems. This study investigated the impact of ISO/IEC 17025 accreditation on the performance of an NMRA Quality Control laboratory to provide evidence of improved quality compliance within a low-resource setting. Pre- and post- accreditation audits of nonconformities for management and technical requirements of the ISO/IEC17025:2005 standards were evaluated from a Quality Control laboratory in the National Agency for Food and Drug Administration and Control (NAFDAC), located in Nigeria, West Africa. The following research questions were addressed: "does accreditation impact the adherence to quality standards?" and "does accreditation decrease the severity of nonconformities in Quality Control laboratory audits?" Statistical analysis of the pre- to post- accreditation audits from the years 2013 through 2017 revealed a significant decrease in the total number of nonconformities (χ2 = 74, p-value = 9.99e-05, r = 0.67). Further examination of audits from the years 2013 through 2018 audits also revealed a reduction in the number of nonconformities (χ2 = 53, p-value = 9.99e-05, r = 0.62). A reduction in the number of major observations and a decrease in the severity of nonconformities was also observed. A higher level of quality compliance was exhibited for the laboratory during the post-accreditation years. Overall, ISO/IEC 17025 accreditation of the NMRA Quality Control laboratory resulted in improved reliability of test reports and enhancement of the laboratory quality system.


A chemical approach to stem-cell biology and regenerative medicine.
An improved understanding of stem-cell and regenerative biology, as well as a better control of stem-cell fate, is likely to produce treatments for many devastating diseases and injuries. Chemical approaches are starting to have an increasingly important role in this young field. Attention has focused on chemical approaches that allow the precise manipulation of cells in vitro to obtain homogeneous cell types for cell-based therapies. Another promising approach is the development of conventional chemical and biological therapeutics to stimulate endogenous cells to regenerate. Such therapeutics can act on target cells or their niches in vivo to promote cell survival, proliferation, differentiation, reprogramming and homing.


Association between positive psychological traits and changes in dietary behaviour related to first COVID-19 lockdown: A general population-based study.
The spread of the coronavirus disease (COVID-19) led many countries to implement lockdown measures, which resulted in changes in dietary behaviours that could persist over the long term and have associated health consequences. Psychological traits may impact these changes given their known association with dietary behaviours. We aimed to investigate in a population-based study, whether positive psychological traits were associated with changes of snacking behaviour and food consumption observed during the first COVID-19 lockdown period. In 2016, levels of optimism, resilience, self-esteem, satisfaction with life, mindfulness and mastery were assessed in 33,766 adults of the French NutriNet-Santé cohort. Snacking and food group consumption were assessed in April-May 2020. Association between psychological traits and changes (no change, increase, decrease) in snacking and food group consumption were assessed using logistic regressions. Multiple correspondence analysis followed by ascending hierarchical classification were used to derive clusters of dietary behaviours. Covariance analyses were used to compare mean scores of psychological traits between clusters. Analyses were adjusted for sociodemographic and lifestyle characteristics, anxiety and depressive symptomatology. Participants with higher levels of optimism, resilience, self-esteem, satisfaction with life, mindfulness or mastery were less likely to change their snacking behaviour and food group consumption of various food groups. Individuals with lower levels were more likely to make changes, with either unhealthy (e.g., less fruits and vegetables, more processed meat) or healthy (e.g., more pasta/rice (whole-grain)) changes. Overall, individuals showed higher levels of positive psychological traits in the "no change" cluster, followed by the "healthy" and the "unhealthy" cluster (all P < 0.05). Individuals with higher levels of optimism, resilience, self-esteem, satisfaction with life, mindfulness or mastery were less impacted by the lockdown in terms of dietary behaviours.


Osteoprotegerin and RANKL regulate bone resorption, density, geometry and strength.
Osteoprotegerin (OPG) and receptor activator of nuclear factor-kappaB ligand (RANKL) are dominant regulators of bone resorption. Many hormones, cytokines and growth factors mediate bone resorption by altering the ratio of RANKL to OPG. RANKL and OPG expression is also altered in numerous bone diseases, and these changes can reflect disease etiology or compensatory responses to disease. RANKL stimulates osteoclast formation, function and survival, and each of these effects is inhibited by OPG. OPG suppresses bone resorption and increases the density, area and strength of both cancellous and cortical bone. Denosumab (AMG 162), a fully human monoclonal antibody to RANKL, shares the pharmacologic attributes of OPG but has a significantly longer half-life that allows less frequent administration.


Differential effects of vigabatrin and zonisamide on the neuropeptide Y system in the hippocampus of seizure prone gerbil.
Changed neuropeptide Y (NPY) system in the hippocampus has been reported in various experimental epileptic models. However, there have been little data concerning the alteration in the NPY system in the epileptic hippocampus following treatment of anti-epileptic drugs (AEDs). In the present study, therefore, we performed analyses of effects of vigabatrin (VGB) and zonisamide (ZNS) treatment on the NPY system in the hippocampus of the seizure sensitive (SS) gerbils. In SS gerbil, NPY immunoreactivity in the hippocampus was lower than that in seizure resistant gerbil. Following VGB treatment, the number of NPY immunoreactive neurons and NPY mRNA expression were increased in the hilus and the hippocampus proper. In contrast, ZNS treatment markedly elevated only the density of NPY immunoreactive fibers in the dentate gyrus, not in the hippocampus proper, as compared with saline-treated animals. These patterns were observed in the dose-dependent manners. These findings suggest that AEDs treatments may distinctly affect the NPY system in the SS gerbil hippocampus.


Production of bioactive triterpenes by Eriobotrya japonica calli.
Callus tissue cultures induced from an axenic leaf of Eriobotrya japonica (Rosaceae) produced triterpenes in large amounts (ca. 50 mg/g dry wt). Nine triterpenes were characterized as ursolic acid, oleanolic acid, 2alpha-hydoxyursolic acid, maslinic acid, tormentic acid, 2alpha, 19alpha-dihydroxy-3-oxo-urs-12-en-28-oic acid, hyptadienic acid and a mixture of 3-O-cis-p-coumaroyltormentic acid and 3-O-trans-p-coumaroyltormentic acid. The triterpene composition in the callus tissues was noticeably different from that in intact leaves. The contents of tormentic acid with antidiabetic action, and 2alpha, 19alpha-dihydroxy-3-oxo-urs-12-en-28-oic acid with anti-HIV activity, were much larger than those in the intact leaves. All of the triterpenes isolated from the callus tissues showed an inhibitory effect comparable to (-)-epigallocatechin gallate (EGCG) of green tea on the activation of Epstein-Barr virus early antigen (EBV-EA) induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). 2alpha, 19alpha-Dihydroxy-3-oxo-urs-12-en-28-oic acid was the most potent inhibitor among them and caused a significant delay of two-stage carcinogenesis on mouse skin.


Analysis of Eight Types of Plant-based Milk Alternatives from the United States Market for Target Minerals and Trace Elements.
A wide variety of commercial plant-based foods that are marketed and sold as alternatives for milk (plant-based milk alternatives or PBMAs) are available to consumers. In this study, PBMAs from the United States (n=85) were subjected to analysis for target minerals (magnesium, phosphorus, selenium, and zinc) to compare their variability across PBMA types, brands, and production lots. Samples were also screened for the environmental contaminant elements arsenic, cadmium, and lead. The eight PBMA types sampled were produced from almond, cashew, coconut, hemp, oat, pea, rice, and soy. Elemental analysis was conducted using microwave-assisted acid digestion followed by inductively coupled plasma-mass spectrometry. The results showed that pea PBMAs contained the highest mean amounts of phosphorus, selenium, and zinc, while soy PBMAs were highest in magnesium. Mean amounts of minerals were lower than those found in milk for the majority of PBMA types. There was significant variation (P<0.05) in amounts of minerals across the majority of product brands. The amounts of phosphorus and magnesium varied across production lots (P<0.05), but the absolute value of these differences was low. Total arsenic was highest in rice PBMAs; amounts of cadmium and lead across PBMAs were generally found at low or non-quantifiable amounts. These results underscore the importance of generating analytical data on the elemental composition of products within the rapidly growing category of PBMA.


Changes in protein synthesis and breakdown rates and responsiveness to growth factors with ageing in human lung fibroblasts.
The effects of insulin, epidermal growth factor, insulin-like growth factor-2 and fetal bovine serum on protein synthesis and protein breakdown have been measured in mid-passage and senescent cultures of human diploid lung fibroblasts. Each of the individual growth factors was a potent inhibitor of protein breakdown with no difference in either the maximum response or sensitivity evident between senescent cells and mid-passage cultures. Binding of 125I-labelled epidermal growth factor per mg of cell protein similarly showed no difference between senescent and confluent mid-passage fibroblasts, although sparse mid-passage cells bound more of the ligand. These results indicate normal binding and normal responsiveness to growth factors in senescent cultures. However, rates of protein synthesis are higher in confluent mid-passage cells and especially so in sparse mid-passage cells than in senescent cells of intermediate density. Furthermore, senescent cells differ from either growth state of mid-passage cells because protein synthesis in aged cells is much less responsive to any of the growth factors or to fetal bovine serum. It is suggested from these results that the reduced ability of serum or growth factors to initiate DNA synthesis or growth in ageing cells may be a consequence of an unresponsive protein synthesis pathway rather than a generalised defect in growth factor action.


Cellular electrophysiological effects of botulinum toxin A on neonatal rat cardiomyocytes and on cardiomyocytes derived from human-induced pluripotent stem cells.
Botulinum toxin A is a well-known neurotransmitter inhibitor with a wide range of applications in modern medicine. Recently, botulinum toxin A preparations have been used in clinical trials to suppress cardiac arrhythmias, especially in the postoperative period. Its antiarrhythmic action is associated with inhibition of the nervous system of the heart, but its direct effect on heart tissue remains unclear. Accordingly, we investigate the effect of botulinum toxin A on isolated cardiac cells and on layers of cardiac cells capable of conducting excitation. Cardiomyocytes of neonatal rat pups and human cardiomyocytes obtained through cell reprogramming were used. A patch-clamp study showed that botulinum toxin A inhibited fast sodium currents and L-type calcium currents in a dose-dependent manner, with no apparent effect on potassium currents. Optical mapping showed that in the presence of botulinum toxin A, the propagation of the excitation wave in the layer of cardiac cells slows down sharply, conduction at high concentrations becomes chaotic, but reentry waves do not form. The combination of botulinum toxin A with a preparation of chitosan showed a stronger inhibitory effect by an order of magnitude. Further, the inhibitory effect of botulinum toxin A is not permanent and disappears after 12 days of cell culture in a botulinum toxin A-free medium. The main conclusion of the work is that the antiarrhythmic effect of botulinum toxin A found in clinical studies is associated not only with depression of the nervous system but also with a direct effect on heart tissue. Moreover, the combination of botulinum toxin A and chitosan reduces the effective dose of botulinum toxin A.


A comparison of direct and indirect video endoscopy.
The advent of microchip video cameras has opened another technology to endoscopy of the gastrointestinal tract: video endoscopy. With direct video endoscopy, the microchip camera is mounted on the distal tip of the endoscope; with indirect video endoscopy, a miniature camera is coupled to the eyepiece of a standard fiberoptic endoscope. Both systems view the procedure on a monitor. The systems were "bench" tested in an electronics laboratory to objectively compare resolution, brightness (luminosity), and color intensity (chroma). Clinical comparison was performed in 619 patients. Both video endoscopy systems provided significant advantages over fiberoptic endoscopes in therapeutic procedures, teaching, and documentation. The direct video system provided the best overall video image; the indirect system had a slightly diminished resolution and luminosity in both the "bench" test and clinical evaluation.


Tumors in F344 rats by oral administration of 1,3-diethyltriazene.
A group of 20 male F344 rats was given by gavage a solution of 2.5 mg of 1,3-diethyltriazene in 0.2 ml corn oil twice a week for 20 weeks. Diethyltriazene is very unstable in hydroxylic solvents, but when administered in oil by gavage it induced tumors in almost all of the treated animals, 16 of which had neoplasms of the forestomach; 13 of which were carcinomas. In addition 9 rats had adenomas of the nasal mucosa, two had alveolar-bronchiolar adenomas of the lung, one had a carcinoma of the esophagus and one an adenocarcinoma of the ileum. Half of the rats had died by week 56 of the experiment, the first at week 40 and the last at week 73. The induction of tumors at sites distant from the stomach indicate that diethyltriazene is more stable in the body than would be predicted from its chemical behavior. Although diethyltriazene is a protected alkyldiazonium ion, its carcinogenic effects in rats differ from ethylnitrosoureas, which are also ethylating agents.


ENT manifestations of alkaptonuria: report on a case series.
Alkaptonuria is an inborn error of metabolism. It is a multisystem disease with characteristic ENT manifestations. This paper reports, for the first time, the ENT findings in a cohort of alkaptonuria patients. Patients attending the National Centre for Alkaptonuria (Royal Liverpool and Broadgreen University Hospitals NHS Trust) underwent a full ENT assessment. Eighteen of the 20 patients (90 per cent) had an ENT sign or symptom. These included discolouration of the pinna, cerumen, nasal septum and pharynx. Discolouration of cerumen may occur before 30 years of age and may therefore be an important early clinical sign. Further audiological assessment of patients is needed to clarify if an association exists between alkaptonuria and hearing loss. Alkaptonuria is a condition that could present to the otolaryngologist. Successful early diagnosis and referral to a specialist centre is essential so that patients can be offered disease-modifying therapy.


Peri-operative physiotherapy improves outcomes for women undergoing incontinence and or prolapse surgery: results of a randomised controlled trial.
Urinary incontinence and pelvic organ prolapse are common complaints in women. Physiotherapy and surgery to correct these conditions are often seen as mutually exclusive. No study has yet investigated their synergistic potential. This study aimed to investigate the role of peri-operative physiotherapy in women undergoing corrective surgery for pelvic organ prolapse and/or incontinence. In this randomised controlled trial, 30 women underwent preoperative physiotherapy and 30 others had no physiotherapy prior to their incontinence and or prolapse surgeries. Comparison was performed on the basis of the following tests: paper towel test, urinary symptom specific health and quality of life questionnaire, frequency/volume chart and pelvic floor muscle manometry. Women were followed up for 3 months. Both groups showed improvement in urinary continence. Significant group differences were noted in the quality of life questionnaire (P= 0.004), urinary symptoms (P= 0.017) and maximum pelvic floor muscle squeeze on manometry (P= 0.022). Diurnal frequency analysis indicates that there is a significant difference in favour of the treatment group (P= 0.024). Routine pre and post operative physiotherapy interventions improve physical outcomes and quality of life in women undergoing corrective surgery for urinary incontinence and or pelvic organ prolapse.


Light-energy conversion in engineered microorganisms.
Increasing interest in renewable resources by the energy and chemical industries has spurred new technologies both to capture solar energy and to develop biologically derived chemical feedstocks and fuels. Advances in molecular biology and metabolic engineering have provided new insights and techniques for increasing biomass and biohydrogen production, and recent efforts in synthetic biology have demonstrated that complex regulatory and metabolic networks can be designed and engineered in microorganisms. Here, we explore how light-driven processes may be incorporated into nonphotosynthetic microbes to boost metabolic capacity for the production of industrial and fine chemicals. Progress towards the introduction of light-driven proton pumping or anoxygenic photosynthesis into Escherichia coli to increase the efficiency of metabolically-engineered biosynthetic pathways is highlighted.


Flaxseed supplementation and early markers of colon carcinogenesis.
Since flaxseed ingestion produces potentially anticarcinogenic lignans in the colon, this study determined whether flaxseed decreases the risk for colon carcinogenesis. Following a single injection of azoxymethane (15 mg/kg body wt.), five groups of male Sprague-Dawley rats were fed a high-fat (20% corn oil) basal diet with or without supplementation with 5% or 10% flaxseed meal (FM) or flaxseed flour (FF) for four weeks. Upon sacrifice, colons were examined for aberrant morphology and cell proliferation. In the descending colon of supplemented groups, the total number of aberrant crypts and foci were significantly reduced by 41-53% and 48-57%, respectively. The labeling index (LI) was also 10-22% lower in these groups, except for the 5% FM group. While these effects are not linearly related to the level of flaxseed fed, it suggests that flaxseed feeding may reduce the risk for colon carcinogenesis.


Accuracy and reproducibility of the assessment of the global ejection fraction using 195mAu and a single-crystal digital gamma camera: influence of collimator design.
A sequence of RAO first-pass studies (first with 99mTc and then twice with 195mAu) was performed in 18 normal volunteers and in 12 CAD patients using two different types of collimator for medium energy: a standard collimator (MEMS) and a special high-sensitivity collimator (MEHS). The following data were compared: the peak count rate, the net LV end-diastolic counts and the LVEF. Using MEMS the end-diastolic counts acquired were so low (12% of 99mTc average value) that EF standard deviation had a mean value of 0.061 (range 0.045-0.081). With MEHS the following results were obtained: 1. the peak count rate and LV net end-diastolic counts with 195mAu were 55% and 50% respectively, of 99mTc values; 2. a good correlation was shown between LVEF values either with 99mTc and 195mAu (r = .97), or with 195mAu sequential studies (r = .98).


Safety of outpatient thyroidectomy: Review of the American College of Surgeons National Surgical Quality Improvement Program.
To investigate national trends in admission status after thyroidectomy in the United States and to evaluate the factors associated with 30-day unplanned readmission and reoperation. Retrospective review of American College of Surgeons National Surgical Quality Improvement Program (ACS-NSQIP) METHODS: The ACS-NSQIP database was queried for patients who underwent a partial or total thyroidectomy between 2005 and 2014. Outpatient surgery was defined as discharge on the day of surgery. Patient demographic information, unplanned hospital readmission, and reoperation were reviewed. Risk factors were identified using logistic regression modeling. A total of 76,604 cases met inclusion criteria as described above. There were 1,473 (1.9%) patients who underwent reoperation and 477 unplanned 30-day readmissions (1.4%) for procedures performed since 2012. There was a significant positive trend in the percentage of thyroidectomy (partial and total) patients who underwent outpatient procedures by year of operation (P < .001). Outpatient procedures were not more likely to have unplanned readmissions or reoperations. Independent patient risk factors for unplanned readmission and reoperation included current dialysis, chronic steroid use, unintentional weight loss, American Society of Anesthesiologists class 3 to 4, and active bleeding disorders. Over the past decade there has been a clear trend toward increasing outpatient thyroid surgery. Thyroidectomy performed as an outpatient was not found to be an independent risk factor for readmission or reoperation. Patients with serious medical comorbidities and active bleeding disorders are at increased risk of unplanned readmission or reoperation and should have their surgery performed on an inpatient basis. 2c. Laryngoscope, 128:1249-1254, 2018.


Improving regional care in the last year of life by setting up a pragmatic evidence-based Plan-Do-Study-Act cycle: results from a cross-sectional survey.
To set up a pragmatic Plan-Do-Study-Act cycle by analysing patient experiences and determinants of satisfaction with care in the last year of life. Cross-sectional postbereavement survey. Regional health services research and development structure representing all health and social care providers involved in the last year of life in Cologne, a city with 1 million inhabitants in Germany. 351 bereaved relatives of adult decedents, representative for age and gender, accidental and suspicious deaths excluded. For the majority (89%) of patients, home was the main place of care during their last year of life. Nevertheless, 91% of patients had at least one hospital admission and 42% died in hospital. Only 60% of informants reported that the decedent had been told that the disease was leading to death. Hospital physicians broke the news most often (58%), with their communication style often (30%) being rated as 'not sensitive'. Informants indicated highly positive experiences with care provided by hospices (89% 'good') and specialist palliative home care teams (87% 'good'). This proportion dropped to 41% for acute care hospitals, this rating being determined by the feeling of not being treated with respect and dignity (OR=23.80, 95% CI 7.503 to 75.498) and the impression that hospitals did not work well together with other services (OR=8.37, 95% CI 2.141 to 32.71). Following those data, our regional priority for action now is improvement of care in acute hospitals, with two new projects starting, first, how to recognise and communicate a limited life span, and second, how to improve care during the dying phase. Results and further improvement projects will be discussed in a working group with the city of Cologne, and repeating this survey in 2 years will be able to measure regional achievements. DRKS00011925.


[Changes in the ionic composition and shape of erythrocytes in response to cholera toxin].
Blood of 10 inbred rats has been investigated after effect of cholera toxin for 1 h by means of roentgenospectral energetic spectrometer Revex-5100 (USA). Simultaneously, erythrocytes have been studied by means of the scanning electron microscope. Contents of sodium, phosphorus, chlorine, serum, potassium, and ferrum ions have been estimated. The cholera toxin changes the contents of the ions mentioned in erythrocytes and contributes to certain changes of their form. A suggestion is made that the cholera toxin changes permeability of the erythrocytes membranes without participation of the cell cyclase systems.


Synthesis, antibacterial properties and bioinformatics computational analyses of novel 8-hydroxyquinoline derivatives.
New heterocyclic derivatives of 8-hydroxyquinoline were prepared and screened as antimicrobial agents. Chemical structures were elucidated and confirmed using different spectroscopic methods such as elemental analysis data, Infrared, Nuclear Magnetic Resonance Spectroscopy. In order to explore their potential biological activity, the "in vitro" antibacterial activity was investigated against [E. coli (ATCC35218), S. aureus (ATCC29213), V. parahaemolyticus (ATCC17802), and P. aeruginosa (ATCC27853)]. The studied compounds exhibited a remarkable antibacterial activity superior to the standard antibiotic (Penicillin G). These new heterocyclic derivatives of 8-hydroxyquinoline, which proved to be potentially effective, can be used as alternative chemical antimicrobial agents applications. It was very interesting to observe that POM (Petra/Osiris/Molinspiration) bioinformatic analyses of the 8-hydroxyquinoline derivative (5) exhibited more important antibacterial activity (MIC = 10-6 mg/mL against V.p and S.a bacteria) and good drug score (DS = 0.71) when compared with Penicillin (DS = 0.33; MIC = 10-3 mg/mL).


A novel human laboratory model for screening medications for alcohol use disorder.
Alcohol use disorder (AUD) is a highly prevalent, chronic relapsing disorder with a high disease burden in the USA. Pharmacotherapy is a promising treatment method for AUD; however, the few FDA-approved medications are only modestly effective. Medications development for AUD is a high priority research area, but the cumbersome drug development process hinders many potential compounds from reaching approval. One area with major opportunities for improvement is the process of screening novel compounds for initial efficacy, also known as early phase 2 trials. Early phase 2 trials incorporate human laboratory paradigms to assess relevant clinical constructs, such as craving and subjective responses to alcohol. However, these controlled paradigms often lack the ecological validity of clinical trials. Therefore, early phase 2 trials can be more efficient and clinically meaningful if they combine the internal validity of experimental laboratory testing with the external validity of clinical trials. To that end, the current study aims to develop and validate a novel early efficacy paradigm, informed by smoking cessation literature, to screen novel medications for AUD. As an established AUD medication, naltrexone will serve as an active control to test both the practice quit attempt model and the efficacy of a promising AUD pharmacotherapy, varenicline. Individuals with current AUD reporting intrinsic motivation to change their drinking will complete a week-long "practice quit attempt" while on study medication. Participants are randomized and blinded to either naltrexone, varenicline, or placebo. During the practice quit attempt, participants will complete daily visits over the phone and fill out online questionnaires regarding their drinking, alcohol craving, and mood. Additionally, participants will undergo two alcohol cue-reactivity sessions. The successful completion of this study will advance medications development by proposing and validating a novel early efficacy model for screening AUD pharmacotherapies, which in turn can serve as an efficient strategy for making go/no-go decisions as to whether to proceed with clinical trials. ClinicalTrials.gov NCT04249882 . Registered on 31 January 2020.


Mitochondrial damage induced by conditions of oxidative stress.
Up to 2% of the oxygen consumed by the mitochondrial respiratory chain undergoes one electron reduction, typically by the semiquinone form of coenzyme Q, to generate the superoxide radical, and subsequently other reactive oxygen species such as hydrogen peroxide and the hydroxyl radical. Under conditions in which mitochondrial generation of reactive oxygen species is increased (such as in the presence of Ca2+ ions or when the mitochondrial antioxidant defense mechanisms are compromised), these reactive oxygen species may lead to irreversible damage of mitochondrial DNA, membrane lipids and proteins, resulting in mitochondrial dysfunction and ultimately cell death. The nature of this damage and the cellular conditions in which it occurs are discussed in this review article.


Fractionation of depectinated sugar beet pulp into cellulose, hemicellulose, and lignin with NaOH/urea/H2O and ionic liquid.
This study proposes a novel and feasible dissolution and fractionation method of depectinated sugar beet pulp (SBP) in NaOH/Urea/H2O, ionic liquid (IL) and alkaline treatment processes. Interestingly, the complicated structure of SBP can be treated with 30 % H2SO4 to increase the dissolution rate. Scanning electron microscope (SEM) analysis confirmed that the appearance of cellulose and hemicellulose obtained by two methods were different. At the same time, two lignin fractions showed irregular high-density clusters, which were composed of a large number of submicron particles. The crystal structure of two cellulose fractions changed from cellulose I to cellulose II. The thermal stability of cellulose and lignin obtained by ionic liquid was slightly better than that obtained by NaOH/urea/H2O. Results of Fourier transform infrared (FTIR) and 13C NMR showed that the chemical structures of SBP cellulose, hemicellulose and lignin regenerated from NaOH/urea/H2O and ionic liquid were similar.


Four glycosides from the leaves of Abeliophyllum distichum with inhibitory effects on angiotensin converting enzyme.
The bioassay-guided fractionation of the n-BuOH extract of Abeliophyllum distichum afforded acteoside (1), isoacteoside (2), rutin (3), and hirsutrin (4). Compounds 1-3 moderately inhibited the angiotensin I converting enzyme activity in a dose-dependent manner. Compounds 1-3 showed the 50% inhibitory concentration values of 228 micro g/mL, 290 micro g/mL, and 278 micro g/mL, respectively.