Reduction of exercise-induced ventricular arrhythmias in mild symptomatic heart failure by benazepril.
We studied 14 patients who had exercise-induced ventricular arrhythmias after a previous Q-wave myocardial infarction. All had symptomatic mild heart failure in New York Heart Association class II and a maximal oxygen consumption between 16 and 20 ml/kg/min. They were treated with the angiotensin converting enzyme inhibitor benazepril (20 mg) and hydrochlorothiazide (50 mg) for 3 months in a double-blind randomized cross-over study. Benazepril improved the maximal oxygen uptake by 15% and exercise time by 18%. Hydrochlorothiazide slightly increased exercise time (5%) and the respiratory exchange ratio but not oxygen consumption. The arrhythmias were nonsustained and reproducible in two baseline recordings. Compared with baseline, benazepril reduced the mean number (3.5 +/- 2.5) (+/- SD) of episodes of ventricular tachycardia by 66%, and total (47.4 +/- 40.9) and paired (5.2 +/- 4.5) premature ventricular contractions by 61% and 62%, respectively. Hydrochlorothiazide did not reduce the number of arrhythmias. Thus an improved cardiac function induced by benazepril is associated with a reduction in exercise-induced ventricular arrhythmias in patients with symptomatic mild heart failure after infarction.


Managing acute chest syndrome of sickle cell disease in an African setting.
Despite the fact that acute chest syndrome contributes immensely to morbidity and mortality in patients with sickle cell anaemia, its exact aetiopathogenesis is very complex and not yet well understood. Therefore, a high index of suspicion is needed in its diagnosis, and appropriate treatment should be commenced as soon as possible to prevent lethal complications of this condition, especially in Nigeria where appropriate diagnostic and therapeutic facilities may not be readily available. This is very important, as it may even develop on hospital admission. There is a need to further investigate preventive measures such as the use of hydroxyurea and the newly introduced Nicosan, especially in those people with recurrent disease, in order to reduce both short- and long-term complications of this syndrome among sickle cell patients in Nigeria.


One hundred sixteen cases of acute liver failure treated with MARS.
The various definitions of acute liver failure do not accurately reflect the differences in clinical signs and prognosis. Liver support devices to improve the clinical condition before liver transplantation (LT) were used in 13 patients with primary nonfunction, 24 with fulminant hepatitis, 17 were affected by delayed nonfunction, and 56 of acute on chronic hepatic failure. The average age of these patients was 41.8 years. The average number of applications of molecular absorbing recirculating system (MARS) was about 6 (range: 1-24). The mean length of application was about 9 hours (range: 8-20). MARS treatment was carried out in HLF patients with continuous acute-on-chronic hepatic failure dialisate flow similar to continuous veno venus hemofiltration (CVVH), albumin flow < 20% of hematic flow, heparin 5/10 UI/kg. In acute on chronic hepatic failure (AoCHF) patients, 6- to 11-hour (average 8.5) treatments were performed for a minimum of three treatments. The majority of patients were treated in the intensive care unit (ICU). Laboratory results were also monitored and showed progressive modification: bilirubin (before treatment 22.37 +/- 11.6 mg/dL, after treatment 11.36 +/- 7.5 mg/dL) and ammonium (before treatment 238.2 +/- 19 microg/dL, after treatment 115.4 +/- 12 microg/dL) showed significant change (P < .01). Lactates (before treatment 3.48 +/- 1.3 mmol/L, after treatment 1.76 +/- 1.1 mmol/L) and creatinine (before treatment 2.36 +/- 0.18 mg/dL, after treatment 1.26 +/- 0.67 mg/dL) also showed significant changes (P < .02 and P < .04). Glasgow Coma Score (GCS) went from 8.6 +/- 1.4 to 11.9 +/- 3.9 (P < .05). The mean middle cerebral artery flow (V media) went from 46 cm/s/26-59) to 73 cm/s (52-106) representing decreased cerebral edema, a difference that was not significant. INR scores (before treatment 2.4 after treatment 1.8) also showed no significant change. The MARS can be applied with tolerability for long periods for patients with PDF and FH as a bridge to transplant. In patients with PDF, it is used for a waiting recovery of the transplanted organ. Therefore MARS can also limit the necessity to perform further transplants.


[Analysis of the genetic background and mechanisms of dissemination of resistance to tetracycline of the Polish population of Campylobacter strains isolated in Poland in years 2007-2008].
In Poland, constant rise of number of Campylobacter strains resistant to tetracycline is observed in Poland. Analysis of the resistant strains showed their strong diversity, including both the different levels of resistance to this drug, large differences in the sequence of the resistant gene tetO, and diverse phylogenetic origin. The study also confirmed the important role of horizontal spread of resistance which, in the event of such a large diversity of resistant strains, can cause further very rapid escalation of resistance of Campylobacter to tetracycline.


Trends and challenges in the valorization of kitchen waste to polyhydroxyalkanoates.
Kitchen waste (KW) is frequently available for free or with a negative cost due to its huge production. It contains a large proportion of organic substances, especially fermentable sugars, which can be used for bioplastic (polyhydroxyalkanoates or PHA) synthesis. Nevertheless, due to the difficulties in processing, various pre-treatments of KW are being investigated to enhance the concentration of simple sugars released during its hydrolysis. The effective use of KW will help in minimizing the issues of its inappropriate disposal. However, the review on KW to bioplastic synthesis is rarely reported in the literature. Hence, this particular review provides a comprehensive summary of the updated research developments in KW valorization and its potency as a feedstock for PHAs synthesis. Additionally, the impacts of KW, its availability, the necessary pre-treatments for the biopolymerization process, as well as the prospects and challenges for industrially generating sustainable PHAs, are critically discussed.


Expedient discovery of novel oxime ester derivatives of piperine/piperine analogs as potent pesticide candidates and their mode of action against Tetranychus cinnabarinus Boisduval.
Nowadays, pest infestation and resistance have appeared as a consequence of repeated and extensive use of pesticides. Thus, development of new effective pesticide candidates in crop protection is highly desirable. Herein, a series of new piperine derivatives containing oxime ester scaffolds were regioselectively and stereoselectively prepared as pesticidal agents. Steric configurations of compounds 2, 5z and 13e were definitively determined by single-crystal X-ray diffraction. Against Tetranychus cinnabarinus, notably, compounds 5f [median lethal concentration (LC50 ) = 0.14 mg mL-1 ] and 5v (LC50 = 0.13 mg mL-1 ) showed >107-fold greater acaricidal activity than piperine (LC50 = 15.02 mg mL-1 ), which were comparable to the commercial acaricide spirodiclofen. Against Aphis citricola, compound 5d (LD50 = 19.12 ng aphid-1 ) exhibited 6.1-fold more potent aphicidal activity than piperine (LD50 = 116.06 ng aphid-1 ). Additionally, through scanning electron microscopy, the toxicology study suggested that the acaricidal activity of piperine derivatives may be related to damage of the cuticle layer crest of T. cinnabarinus. The structure-activity relationships suggested that 3,4-dioxymethylene of piperine was crucial for its acaricidal activity; and introduction of a certain length of aliphatic chain at the C-2 position was beneficial to the aphicidal and acaricidal activities. Compounds 5f and 5v are potential leads for further structural modification as acaricidal agents. © 2023 Society of Chemical Industry.


Effect of platelet inhibitors on thromboembolism after implantation of a Novacor N100--preliminary results.
Left-ventricular assist device implantation (LVAD) is still associated with thromboembolism as the optimal anticoagulation is still unclear. We report on the effects of adding platelet inhibitors to our anticoagulation regimen in our Novacor LVAD program. Oral platelet aggregation inhibitors (aspirin 330 mg + dipyridamole 75 mg, three times per day) were added to the heparin/phenprocoumon treatment in 9 patients starting on postoperative day 3 to 7 (group A). Of the previous 41 patients, the last 20 patients served as a control group (group B), to reduce any learning curve effect. The mean interval of mechanical support between the two groups was comparable (group A vs B: 148 +/- 127 vs 104 +/- 61 days, n.s.). Accordingly, the cumulative support was much lower in group A (1051 days) as compared to group B (2091 days). In group B, 10 patients (50%) developed clinically evident thromboembolism. The number of events ranged from 1 to 10 (mean 1.4 +/- 2.3), with a total of 32. With addition of platelet inhibitors, the incidence of cerebral embolism dramatically dropped, as only one patient presented with transient ischemic attacks in group A (p < 0.05). Thoracic bleeding as defined by excessive drainage losses requiring redo thoracotomy did not increase (group A vs B: 22% vs 20%, n.s.). Addition of platelet inhibitors to heparin/phenprocoumon effectively prevents thromboembolism. However, platelet inhibitors should be postponed until sufficient hemostasis is achieved, since too early administration is associated with an increased risk of bleeding.


The exocytosis induced in HL-60 cells by 4-hydroxynonenal, a lipid peroxidation product, is not prevented by reduced glutathione.
The effect of reduced glutathione (GSH) was studied on exocytosis triggered by 4-hydroxynonenal in HL-60 cells induced to differentiate towards the granulocytic cell line by dimethylsulfoxide; we measured beta-glucuronidase secretion from cells incubated at 37 degrees C in the presence of 5 mM GSH. GSH addition to the cell suspensions failed to induce any significant change of the exocytosis stimulated by HNE concentrations between 10(-8) and 10(-6) M. In contrast however, 5 mM GSH was able to fully prevent the release of lactate dehydrogenase observed in the presence of 50 microM HNE, a concentration much higher than that able to stimulate the exocytotic secretion. As the activation of phosphoinositide-specific phospholipase C (PLC) has been shown to play a major role in HNE-induced exocytosis, we studied the GSH effect on the breakdown of phosphatidylinositol-4,5-bisphosphate added to plasma membranes isolated from rat neutrophils and incubated in the presence of increasing concentrations of the aldehyde. In neutrophil membranes HNE induced a significant increase of PLC activity when used in the same concentrations as those able to stimulate beta-glucuronidase secretion in DMSO-differentiated HL-60 cells; the presence of 5 mM GSH failed to prevent its action. Our results suggest that these low aldehyde concentrations, which have actually been found in exudates, may increase tissue damage in inflammation through the release of lytic enzymes by neutrophils; it seems unlikely that their effects could be influenced by the levels of -SH groups present in the exudate and by its protein concentration.


Correction: Huang et al. Deep-Learning Based Label-Free Classification of Activated and Inactivated Neutrophils for Rapid Immune State Monitoring. Sensors 2021, 21, 512.
The authors wish to make the following correction to their paper [...].


Ligand-dependent and -independent processes in B-cell-receptor-mediated signaling.
The B cell antigen receptor (BCR) is a protein complex expressed on the surface of immature and mature B cells. After ligand-induced aggregation, this complex generates signals that lead to a variety of biological outcomes, including survival, proliferation and differentiation. During B cell development intermediate forms of the BCR are expressed on the surface. The composition of these pro- and preBCR complexes reflects the ordered assembly of the BCR complex and they exist to generate signals for positive selection at defined developmental checkpoints. Because these receptors lack the ability to bind conventional ligands, the pro- and preBCR have been postulated to signal via ligand-independent processes. This ligand-independent or constitutive signal may also play a role in the survival of peripheral mature B cells. Here we discuss the evidence for ligand-independent functions for the BCR and postulate how it may be regulated and linked to biological processes associated with B cell development and survival.


Acquired aphasia in children: revisited.
Observations of acquired aphasia in a more numerous series of children with cortical lesions modifies somewhat the conclusions of an earlier study. Aphasic syndromes in these children seem to result almost exclusively from left hemispheric lesions in right-handers. Aphasia is more frequent, at least among the youngest children, than in adults. The factors most responsible for language disorders in these children are lesion localization and etiology.


Effects of salmon oil and corn oil on plasma lipid level and hepato-biliary cholesterol metabolism in rats.
The aim of this work was to compare the effects of n-3 and n-6 fatty acids on plasma lipid level and hepato-biliary cholesterol metabolism by studying rats fed semi-synthetic diets enriched with either 10% salmon oil, 10% corn oil, or a blend of 6% corn oil and 4% salmon oil. After 4 weeks of feeding, a drop in plasma lipid level was noted in the salmon oil group in comparison to the control group, whereas no change was observed in the corn oil group. An increase in production of cholesterol ester by the liver was recorded in the salmon oil group with a marked enhancement in acyl-CoA:cholesterol acyltransferase (ACAT: EC 18.104.22.168) activity and hepatic cholesterol concentration. Corn oil did not affect either ACAT activity or hepatic cholesterol storage. All bile parameters (flow, bile salts, phospholipids, cholesterol) increased in the salmon oil group, but the molar ratio of cholesterol participation in the bile secretion decreased. These changes in bile composition, as well as in hepatic metabolism of cholesterol, may help to explain the hypolipidemia following the intake of fish oil.


Confirmation of nonconvulsive limbic status epilepticus with the sodium amytal test.
Is it a seizure? This question can be difficult for a clinician to answer, and it may be more critical if the possible seizure lasts >30 min. Long-duration questionable seizure activity changes the question to, "Is it status epilepticus?" Status epilepticus (SE) can be divided into convulsive and nonconvulsive types. Convulsive SE is the most easily recognized, whereas nonconvulsive SE is more clinically variable and controversial. The term nonconvulsive SE is more often applied to patients who are severely obtunded or comatose with minimal or no motor movements, or in a stupor of altered consciousness reflecting generalized ictal activity. Nonconvulsive SE also can be caused by focal seizure activity, sometimes restricted to deep small volumes of brain in which scalp EEG may not be diagnostic. We present the case of a patient who had dominant limbic hippocampal SE, but in whom the diagnosis could not be confirmed until a modified novel use of the sodium amytal test was performed.


Expression profile of HSP genes during different seasons in goats (Capra hircus).
The present study has demonstrated the expression of HSP60, HSP70, HSP90, and UBQ in peripheral blood mononuclear cells (PBMCs) during different seasons in three different age groups (Groups I, II, and III with age of 0-2, 2-5, and >5 years, respectively) of goats of tropical and temperate regions. Real-time polymerase chain reaction was applied to investigate mRNA expression of examined factors. Specificity of the desired products was documented using analysis of the melting temperature and high-resolution gel electrophoresis to verify that the transcripts are of the exact molecular size predicted. The mRNA expression of HSP60, HSP90, and UBQ was significantly higher (P < 0.05) in all age groups during peak summer season as compared with peak winter season in both tropical and temperate region goats. HSP70 mRNA expression was significantly higher (P < 0.05) during summer season as compared with winter season in tropical region goats. However, in the temperate region, in goats from all the three age groups studied, a non-significant difference of HSP70 expression between summer and winter seasons was noticed. In conclusion, results demonstrate that (1) HSP genes are expressed in caprine PBMCs and (2) higher expression of HSPs during thermal stress suggest possible involvement of them to ameliorate deleterious effect of thermal stress so as to maintain cellular integrity and homeostasis in goats.


Acute hyperpolarization and elongation of cochlear outer hair cells on superfusion with cis-platinum.
The acute effects of cis-platinum on isolated cochlear outer hair cells (OHC) were investigated with whole-cell patch-clamps and measurements of cell length changes. Our findings demonstrated that cis-platinum reversibly induced a hyperpolarization and cellular elongation. These results suggest that the effects produced are the result of an interaction between cis-platinum and transduction channels in OHC. These acute effects are distinctly different from the chronic, irreversible ones that are followed by death of the OHC. The exact mechanism of these chronic effects remains unknown as yet.


Ultrastructural midgut events in Culicidae larvae fed with Bacillus sphaericus 2297 spore/crystal complex.
Ingestion of Bacillus sphaericus 2297 spore/crystal complex by Culicidae larvae Anopheles stephensi, Culex pipiens subsp. pipiens and Aedes aegypti was rapidly followed by a dissolution of the protein crystalline inclusions inside the anterior stomach of the three species. During the first day of intoxication, B. sphaericus spores germinated within the midgut lumen, and were in a vegetative stage between 36-48 h after ingestion when the larvae began to die. Ultrastructural observations focused on larval midgut showed alterations which differed according to the mosquito species, being localized mainly in the gastric caeca and posterior stomach. With the bacterial concentration used, neither general cell swelling nor complete breakdown of the midgut epithelium was recorded before larval death. In A. stephensi larval midgut epithelium large low-electron-density areas appeared, rough endoplasmic reticula formed numerous concentrical structures and mitochondria swelled. Large vacuoles (of unknown origin) appeared early on in the C. pipiens midgut cells, and rough endoplasmic reticula broke into small vesicles. Midgut epithelial cells of A. aegypti showed mitochondria swelling except in the anterior stomach, and a vacuolisation of smooth reticula: these aspects remained unchanged until the larvae died.


Photothermal combined gene therapy achieved by polyethyleneimine-grafted oxidized mesoporous carbon nanospheres.
Combining controllable photothermal therapy and efficacious gene therapy in a single platform holds great promise in cancer therapy due to the enhanced combined therapeutic effects. Herein, polyethyleneimine-grafted oxidized mesoporous carbon nanospheres (OP) were developed for combined photothermal combined gene therapy in vitro and in vivo. The synthesized OP was characterized to have three dimensional spherical structure with uniformed diameter, ordered mesopores with graphitic domains, high water dispersion with zeta potential of +22 mV, and good biocompatibility. Consequently, OP was exploited as the photothermal convertor with strong NIR absorption and the gene vector via electrostatic interaction, which therefore cannot only deliver the therapeutic gene (pING4) to tumors for gene therapy, but also can eliminate the tumors by photothermal ablation. Moreover, the improved gene therapy accompanied by the NIR photothermally enhanced gene release was also well achieved based on OP. The excellent combined therapeutic effects demonstrated in vitro and in vivo suggested the OP's potential for cancer therapy.


Is methoxydine a new rapid acting antidepressant for the treatment of depression in alcoholics?
Methoxydine is a dissociative anaesthetic belonging to the arylcyclohexylamine class. This substance shows pharmacodynamic similarities with ketamine, a medication with demonstrated rapid-acting antidepressant effects. Like ketamine, results of binding assays have shown that methoxydine is an uncompetitive antagonist of NMDA receptor approximately as potent as ketamine, but less potent than PCP. Furthermore, unlike ketamine, it acts as a dopamine, serotonin, and noradrenaline reuptake inhibitor as well as an agonist at sigma-1, sigma-2, and opioid receptors. The hypothesis is that methoxydine can produce rapid antidepressant effects in depressed patients with high risk of suicide, including depressed alcoholics.


Circulating Tumor Cells are an Independent Risk Factor for Poor Prognosis in Patients with Gallbladder Adenocarcinoma.
The study aimed to evaluate the prognostic impact of circulating tumor cells (CTCs) in patients with gallbladder adenocarcinoma after resection. Between January 2018 and January 2021, 101 consecutive patients with gallbladder adenocarcinoma were included. CTCs were detected and enumerated using the CanPatrol® technique. The follow-up period ended in January 2023. The cancer-specific survival (CSS) and disease-free survival (DFS) were calculated using log-rank and Cox regression analyses. CTCs were detected positively in 61.54% (8/13) of the patients in the non-operation group and 13.64% (12/88) in the operation group. In the operation group, the median CSS for CTCs-positive and CTCs-negative patients was 5.0 and 9.5 months (P < 0.001), respectively, and DFS was 2.8 and 5.0 months at stage III (P < 0.001), respectively. In the non-operation group, the median CSS for CTCs-positive and CTCs-negative patients was 3.5 and 6.5 months (P = 0.0031), respectively. The median CSS for CTCs-positive patients in the operation group was similar to that in the non-operation group (P = 0.67). Multivariate analyses showed that positive CTCs was an independent risk factor for poor CSS (HR 0.066, 95% CI 0.021-0.206, P < 0.001) as well as lymph infiltration (HR 0.320, 95% CI 0.110-0.930, P = 0.036), without R0 curative resection (HR 7.520, 95% CI 2.100-26.931, P = 0.002), and without adjuvant chemotherapy (HR 7.730, 95% CI 2.416-24.731, P < 0.001). Positive CTCs was an independent predictor of poor prognosis after resection in patients with gallbladder adenocarcinoma. Preoperative detection of CTCs may play an important guiding role in formulating treatment strategies for these patients.


A highly specific 125I-radioimmunoassay for cholic acid conjugates.
Several modifications of the immunization procedure permitted development of a highly specific radioimmunoassay (RIA) for cholic acid conjugates. Antiserum was produced in guinea pigs using cholic acid-thyroglobulin complex as immunogen. 125-I-Cholyglycylhistamine was prepared as radioactive ligand according to a modification of the method of Spenney et al. (Spenney, J.G., Johnson, B.J., Hirschowitz, B.I., Mihas, A.A. and Gibson, R. (1977) Gastroenterology 72, 305--311). The association constant of the antisera to taurocholic acid was 1.8 x 10(7) l/mol, the working range of the assay between 9.5--890 pmol. Cross-reactivities of the antiserum to bile acids other than cholic acid species were less than 3%, which is lower than for any published bile acid RIA. Concentrations of cholic acid conjugates in sera obtained from 17 healthy fasting volunteers ranged from 0.4--1.9/mumul/l.


Comparing Caliper versus Computed Tomography Measurements of Cranial Dimensions in Children.
We have found no study assessing the accuracy of caliper cranium measurements compared to computed tomography (CT) measurements of the head. The objective of this study was to assess the reliability of caliper measurements in comparison to CT measurements. Retrospective chart review. This study includes all patients evaluated for head shape abnormality between 2010 and 2019 at a single academic medical center. Eighty-nine patients who had CT head scans were identified, and their caliper measurements of anterior-posterior and transverse head dimensions were documented. There was no statistically significant difference between the CT and caliper measurements. Caliper measurements are a simple and reliable way to assess cranial vault proportionality with growth after cranial vault reconstruction. 4 Laryngoscope, 131:773-775, 2021.


Precursors of taurine in female genital tract: effects on developmental capacity of bovine embryo produced in vitro.
Two precursors of taurine have been studied: cysteamine and hypotaurine. Cysteamine has been quantified in genital secretions and found in follicular fluids of all species tested. On the contrary cysteamine was not detected (or traces) in tubal fluids of the same species. Addition of 50, 100 or 250 microM of cysteamine to the maturation medium used in the culturing of bovine oocytes did not improve the cleavage rate nor the embryo's developmental potential in vitro. Furthermore, at 250 microM, cysteamine seems to be toxic to the embryo. Addition of 0.5-1 mM hypotaurine to the bovine embryo culture medium improved significantly blastocyst production and quality. The respective roles of these 2 taurine precursors on maturation and embryo development are discussed.


A quantitative ultrastructural analysis of changes in hamster cheek-pouch epithelium treated with vitamin A.
The ultrastructural changes induced by the topical application of retinol acetate on hamster cheek pouch epithelium were evaluated using stereological analysis. Electron micrographs were prepared of the basal and superficial regions of the nucleated cell layer of the epithelium obtained from 3 treated and 3 control animals and examined at two levels of magnification. A total of 528 micrographs were analyzed using a coherent double lattice test system. Although the mean thickness of the nucleated cell layer did not change significantly after 10 days of treatment with retinol acetate the formation of keratinized squames was completely inhibited. This was paralleled by significant changes in the volume density of a number of organelles in both the basal and superficial strata. Rough endoplasmic reticulum increased significantly whereas filaments, which maintained a constant diameter of approximately 9 nm, keratohyalin granules and membrane-coating granules decreased in both strata. Desmosomes also showed a significant decrease in numerical area density in the treated tissues. In contrast, no changes were observed in the volume density of the Golgi apparatus, free ribosomes or mitochondria in the treated epithelium. It is concluded that this treatment provides an epithelium lacking all features of keratinization and may be a useful model for examining metabolic activities specifically associated with keratinization.


Estradiol inhibits hyaluronic acid synthase 1 expression in human vascular smooth muscle cells.
Epidemiological and clinical data suggest that estrogen retards the progression of atherosclerosis. This study aims to elucidate whether the phenotypic regulation of human vascular smooth muscle cells (VSMC) by estrogen may involve effects on the hyaluronan matrix. VSMC were synchronized by serum withdrawal and subsequently stimulated with 0.001, 0.01, 0.1 and 1 μM estradiol (E(2)) in the presence or absence of platelet-derived growth factor BB (PDGF-BB) for 24 h. E(2) reduced mRNA-expression of hyaluronic acid synthase (HAS) 1 in the presence and absence of PDGF-BB. In contrast, HAS2- and HAS3-mRNA-expression were not affected. This E(2)-mediated effect on HAS1 mRNA-expression was accompanied by reduced hyaluronan secretion and a shift of HA toward lower molecular weight as evidenced by molecular sieve chromatography. The downregulation of HAS1 was abrogated by the estrogen receptor (ER) α and β antagonist ICI182780 and could be mimicked by the ERα-agonist propyl-pyrazole triol (PPT). On the contrary, the ERβ-agonist diarylpropionitrile (DPN) had no effect on HAS1 mRNA-expression. To investigate whether the downregulation of HAS1 was causally involved in the phenotypic regulation of human VSMC by E(2), lentiviral overexpression of HAS1 was conducted. Overexpression of HAS1 abrogated the inhibition of sustained ERK1/2 phosphorylation and in turn inhibition of DNA-synthesis by E(2). For the first time this study provides strong evidence that HAS1-driven HA-synthesis is a target of E(2) in human VSMC and that E(2) mediates part of its anti-proliferative effects through an ERα-dependent inhibition of HA-synthesis.


Carcinogenic liver fluke Opisthorchis viverrini oxysterols detected by LC-MS/MS survey of soluble fraction parasite extract.
Liquid chromatography in tandem mass spectrometry (LC-MS/MS) has emerged as an informative tool to investigate oxysterols (oxidized derivatives of cholesterol) in helminth parasite associated cancers. Here, we used LC-MS/MS to investigate in soluble extracts of the adult developmental stage of Opisthorchis viverrini from experimentally infected hamsters. Using comparisons with known bile acids and the metabolites of estrogens, the LC-MS data indicated the existence of novel oxysterol derivatives in O. viverrini. Most of these derivatives were ramified at C-17, in similar fashion to bile acids and their conjugated salts. Several were compatible with the presence of an estrogen core, and/or hydroxylation of the steroid aromatic ring A, hydroxylation of both C-2 and C-3 of the steroid ring and further oxidation into an estradiol-2,3-quinone.


The profiles of gamma-H2AX along with ATM/DNA-PKcs activation in the lymphocytes and granulocytes of rat and human blood exposed to gamma rays.
Establishing a rat model suitable for γ-H2AX biodosimeter studies has important implications for dose assessment of internal radionuclide contamination in humans. In this study, γ-H2AX, p-ATM and p-DNA-PKcs foci were enumerated using immunocytofluorescence method, and their protein levels were measured by Western blot in rat blood lymphocytes and granulocytes exposed to γ-rays compared with human blood lymphocytes and granulocytes. It was found that DNA double-strand break repair kinetics and linear dose responses in rat lymphocytes were similar to those observed in the human counterparts. Moreover, radiation induced clear p-ATM and p-DNA-PKcs foci formation and an increase in ratio of co-localization of p-ATM or p-DNA-PKcs with γ-H2AX foci in rat lymphocytes similar to those of human lymphocytes. The level of γ-H2AX protein in irradiated rat and human lymphocytes was significantly reduced by inhibitors of ATM and DNA-PKcs. Surprisingly, unlike human granulocytes, rat granulocytes with DNA-PKcs deficiency displayed a rapid accumulation, but delayed disappearance of γ-H2AX foci with essentially no change from 10 h to 48 h post-irradiation. Furthermore, inhibition of ATM activity in rat granulocytes also decreased radiation-induced γ-H2AX foci formation. In comparison, human granulocytes showed no response to irradiation regarding γ-H2AX, p-ATM or p-DNA-PKcs foci. Importantly, incidence of γ-H2AX foci in lymphocytes after total-body radiation of rats was consistent with that of in vitro irradiation of rat lymphocytes. These findings show that rats are a useful in vivo model for validation of γ-H2AX biodosimetry for dose assessment in humans. ATM and DNA-PKcs participate together in DSB repair in rat lymphocytes similar to that of human lymphocytes. Further, rat granulocytes, which have the characteristic of delayed disappearance of γ-H2AX foci in response to radiation, may be a useful experimental system for biodosimetry studies.


A non-excision uvr-dependent DNA repair pathway of Escherichia coli (involvement of stress proteins).
In UV-irradiated excision-proficient (uvr+) Escherichia coli, pre-induced by simultaneous pre-starvation for thymine (T) and amino acids (AAs), and/or a low UV pre-dose applied after prestarvation for AAs, pyrimidine dimer excision (PDE) is reduced without an adequate increase of UV sensitivity and UV mutagenesis. The unexcised lesions are tolerated by a putative repair pathway that is uvr dependent but does not involve excision. The process consists of PDE inhibition, which requires outer membrane protease OmpT, and subsequent pyrimidine dimer (PD) toleration, which may be mediated by interaction with a sister duplex using a number of SOS and stress-inducible proteins.


Blockade of central GLP-1 receptors prevents CART-induced hypophagia and brain c-Fos expression.
Central administration of both CART and GLP-1 reduces feeding and increases c-Fos in brain areas associated with food intake. To determine whether aspects of CART's effects were mediated through GLP-1's action, we examined whether the GLP-1 receptor antagonist des-His1-Glu9-exendin-4 (EX) blocked CART-induced feeding inhibition, and c-Fos activation. An i.c.v. dose of 100 microg EX blocked the feeding inhibitory action of 1 microg of CART i.c.v. and prevented CART-induced c-Fos expression at multiple hindbrain and hypothalamic sites. These data suggest that i.c.v. CART administration activates a central release of GLP-1 to inhibit feeding and produce widespread neural activation.


Heavy metal pollution in sediments of a typical mariculture zone in South China.
The heavy metal inventory and the ecological risk of the estuarine sediments in Hailing Bay, an important maricultural zone along the southern coast of China, were investigated. Results show that the surface sediments were mainly polluted by As (2.17-20.34 mg/kg), Ni (1.37-42.50mg/kg), Cu (1.21-58.84 mg/kg) and Zn (11.69-219.22 mg/kg). Furthermore, the aquafarming zone was significantly more polluted than the non-aquafarming zone, and cluster analysis suggested additional sources of heavy metal input in the aquafarming zone. As, Cr, Cu, Ni, Pb and Zn were mainly present in the non-bioavailable residual form in the surface sediments, whereas Cd was predominantly in the highly mobile acid soluble and reducible fractions. The ecological risk of the polluted sediments stemmed mainly from Cd, and from As, Cu and Pb to less degrees. The highest potential risks occurred near the aquaculture base, indicating the need to control heavy metal inputs from aquafarming activities.


Cytochrome c oxidase deficient muscle fibres: substantial variation in their proportions within skeletal muscles from patients with mitochondrial myopathy.
Mitochondrial DNA (mtDNA) disease is a common cause of myopathy and the presence of histochemically demonstrated cytochrome c oxidase (COX) deficiency is an extremely useful diagnostic feature. However, there is currently no quantitative information regarding the variability of COX deficiency within or between muscles. This study addresses this issue by studying a number of skeletal muscle samples obtained at post-mortem from three patients with mitochondrial disease due to established mitochondrial DNA defects. COX deficient muscle fibres were enumerated in sections of these muscles and analysed according to patient, individual muscle, position within a particular muscle and sample size. Descriptive statistics were generated followed by an analysis of variance (ANOVA) to assess the effect of these parameters on the mean percentage of COX deficient fibres. We observed statistically significant variation in the percentage of COX deficient fibres within individual muscles from each patient for samples sizes of between 100 and 400 fibres. Our results have implications for the way in which biopsies of skeletal muscle are used for the assessment of disease severity, progression and response to treatment.


Longevity of lobsters is linked to ubiquitous telomerase expression.
Mammals have high growth rates in embryonic and juvenile phases and no growth in adult and senescent phases. We analyzed telomerase activity in a fundamentally different animal which grows indeterminately. Lobsters (Homarus americanus) grow throughout their life and the occurrence of senescence is slow. A modified TRAP assay was developed and the lobster telomeric repeat sequence TTAGG was determined. We detected telomerase activities which were dependent on RNA and protein components, required dGTP, dATP and dTTP, but not dCTP. Telomerase products with a five nucleotide periodicity were generated. High telomerase activities were detected in all lobster organs. We conclude that telomerase activation is a conserved mechanism for maintaining long-term cell proliferation capacity and preventing senescence, not only in cellular models or embryonic life stages but also in adult multicellular organisms.


H-2 restriction specificity of T cells from H-2 incompatible radiation bone marrow chimeras: further evidence for the absence of crucial influence of the host/thymus environment on the generation of H-2 restricted TNP-specific T lymphocyte precursors.
Experiments were conducted to answer the questions related to (a) the role played by the antigen-presenting cells (APCs) present within the thymus and (b) the effect of radiation dose to the recipients on the H-2 restriction profile of TNP-specific cytotoxic T lymphocyte precursors (CTLP) recovered from spleens and/or thymuses of H-2 incompatible radiation bone marrow chimeras (BMC). The H-2 restriction profile of intrathymically differentiating TNP-specific CTLPs was also analyzed in order to test an argument that donor-H-2 restricted CTLP detected in spleens of H-2 incompatible BMC were due to the extrathymically differentiated T cells under the influence of donor-derived lymphoreticular cells. The results indicated the following: (i) splenic T cells from B10(H-2b) leads to (B10(H-2b) leads to B10.BR(H-2k)) chimeras, which were constructed by irradiating primary B10 leads to B10.BR chimeras with 1100 R and reconstituting them with donor-type (B10) bone marrow cells as long as 8 months after their construction, manifested restriction specificities for both donor- and host-type H-2, (ii) splenic T cells from two types of (B10 X B10.BR)F1 leads to B10 chimeras which were reconstituted after exposure of the recipients with either 900 or 1100 R with donor-type bone marrow cells generated both donor- and host-H-2 restricted TNP-specific cytotoxic T cells, and (iii) the TNP-specific CTLPs present in the regenerating thymuses of B10.BR leads to B10 and (B10 X B10.BR)F1 leads to B10 chimeras 4 weeks after their construction were also shown to manifest both donor- and host-H-2 restriction specificities. The significance of these findings on the H-2 restriction profile of CTLP generated in BMCs is discussed.


Angiogenesis and extracellular matrix remodelling in bronchioloalveolar carcinomas: distinctive patterns in mucinous and non-mucinous tumours.
Bronchioloalveolar carcinomas (BACs) are rare primitive lung adenocarcinomas growing along the alveolar septum without stromal, vascular or pleural invasion. We report an immunohistochemical study of their vascular microenvironment. In three mucinous BACs (M-BAC) and three non-mucinous BACs (NM-BAC) we examined the following parameters in comparison with the normal lung: (i) constituents of the alveolar extracellular matrix; (ii) qualitative and quantitative changes of alveolar capillaries; and (iii) expression of vascular endothelial growth factor (VEGF) by tumour cells. In M-BAC, the alveolar matrix was unchanged compared with the normal parenchyma. Capillaries expressed normal alveolar endothelial markers and their average surface was calculated, as in normal lung, as 8%. VEGF was negative in tumour cells. In NM-BAC, the alveolar wall was thickened by deposits of fibronectin and type III collagen containing myofibroblasts and the basement membrane was disrupted. Capillaries did not retain alveolar endothelial markers and their surface was calculated as 19%. Tumour cells expressed high levels of VEGF. In contrast to NM-BAC, M-BAC do not modify the alveolar structure and seem to exploit the normal alveolar vascular bed to grow, without inducing neoangiogenesis. A better understanding of the mechanisms of growth of lung cancers may have implications for future anti-angiogenic therapeutic strategies.


Effects of tryptamine on active sodium and chloride transport in the isolated bullfrog cornea.
The effects of the serotonin analogue, tryptamine, on the active transepithelial transport of Na+ and Cl- in the in vitro bullfrog cornea were studied. Tryptamine, 1 mM, inhibited both the short-circuit current (Isc) and potential difference (PD) of corneas transporting either Na+ alone or both Na+ and Cl-. The electrical resistance, R, increased in all cases. Both unidirectional Na+ and Cl- fluxes were decreased by tryptamine and these changes accounted for the inhibitory effects on the Isc. The effects of tryptamine were considered along with those of 2 mM theophylline and 0.1 mM ouabain. Tryptamine inhibited the Isc and both undirectional Cl- fluxes which were previously stimulated by theophylline. Theophyline addition, after tryptamine preincubation, increases the Cl- undirectional fluxes but did not restore the inhibited Isc. The inhibitory effects of tryptamine on active Na+ and Cl- transport were different from those of ouabain. While both drugs inhibited the forward Na+ and Cl- fluxes, their backfluxes decreased with tryptamine and increased with ouabain. The addition to the bathing solution of tryptamine after ouabain preincubation reduced the ouabain-increased backward Cl- flux and further increased the electrical resistance. These results are analyzed in terms of an electrical model from which it appears that trypamine's mechanism of action was to decrease cellular permeability to the transepithelial movement of Na+ and Cl-.


Hypnotic action of ethyl beta-carboline-3-carboxylate, a benzodiazepine receptor antagonist, in cats.
The present study demonstrates that ethyl beta-carboline-3-carboxylate (beta-CCE), a benzodiazepine receptor antagonist, has hypnotic and sedative actions in cats. Moreover, at a dose that does not by itself affect sleep, beta-CCE reverses the action of diazepam on sleep organization. The hypnotic effect of subcutaneous administration of beta-CCE (5 mg/kg) lasts for 3-4 h. During this period, deep slow wave sleep (deep non-REM sleep) and paradoxical sleep (REM sleep) significantly increase, while wakefulness markedly decreases. These results, which are quite opposite to the effects of benzodiazepines on sleep organization in cats, support the notion that beta-CCE also acts as a benzodiazepine antagonist of sleep organization.


Zooming in on biological processes with fluorescence nanoscopy.
Fluorescence nanoscopy enables the study of biological phenomena at nanometer scale spatial resolution. Recent biological studies using fluorescence nanoscopy have showcased the ability of these techniques to directly observe protein organization, subcellular molecular interactions, structural dynamics, electrical signaling, and diffusion of cytosolic proteins at unprecedented spatial resolution. Super-resolution imaging techniques critically rely on bright fluorescent probes such as organic dyes or fluorescent proteins. Recently, these methods have been extended to live cells and multicolor, three-dimensional imaging, thereby providing exquisite spatiotemporal resolutions of the order of 10-20 nm and 1-2 s for subcellular imaging. Further improvements in image processing algorithms, labeling techniques, correlative microscopy, and development of advanced fluorescent probes will be required to achieve true molecular-scale resolution using these techniques.


Insights into nanomycoremediation: Secretomics and mycogenic biopolymer nanocomposites for heavy metal detoxification.
Our environment thrives on the subtle balance achieved by the forever cyclical nature of building and rebuilding life through natural processes. Fungi, being the evident armor of bioremediation, is the indispensable element of the soil food web, contribute to be the nature's most dynamic arsenal with non-specific enzymes like peroxidase (POX), glutathione peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD), non-enzymatic compounds like thiol (-SH) groups and non-protein compounds such as glutathione (GSH) and metallothionein (MT). Recently, the area of nanomycoremediation has been gaining momentum as a powerful tool for environmental clean-up strategies with its ability to detoxify heavy metals with its unique characteristics to adapt mechanisms such as biosorption, bioconversion, and biodegradation to harmless end products. The insight into the elaborate secretomic processes provides us with huge opportunities for creating a magnificent living bioremediation apparatus. This review discusses the scope and recent advances in the lesser understood area, nanomycoremediation, the state-of-the-art, innovative, cost-effective and promising tool for detoxification of heavy metal pollutants and focuses on the metabolic capabilities and secretomics with nanobiotechnological interventions.


Quantification of sphingosine 1-phosphate by validated LC-MS/MS method revealing strong correlation with apolipoprotein M in plasma but not in serum due to platelet activation during blood coagulation.
Sphingosine 1-phosphate (S1P) is a signalling sphingolipid affecting multiple cellular functions of vascular and immune systems. It circulates at submicromolar levels bound to HDL-associated apolipoprotein M (apoM) or to albumin. S1P in blood is mainly produced by platelets and erythrocytes, making blood sampling for S1P quantification delicate. Standardisation of sampling is thereby of great importance to obtain robust data. By optimising and characterising the extraction procedure and the LC-MS/MS analysis, we have developed and validated a highly specific and sensitive method for S1P quantification. Blood was collected from healthy individuals (n = 15) to evaluate the effects of differential blood sampling on S1P levels. To evaluate correlation between S1P and apoM in different types of plasma and serum, apoM was measured by ELISA. The method showed good accuracy and precision in the range of 0.011 to 0.9 μM with less than 0.07 % carryover. We found that the methanol precipitation used to extract S1P co-extracted apoM and several other HDL-proteins from plasma. The platelet-associated S1P was released during coagulation, thus increasing the S1P concentration to double in serum as compared to that in plasma. Gel filtration chromatography revealed that the platelet-released S1P was mainly bound to albumin. This explains why the strong correlation between S1P and apoM levels in plasma is lost upon the clotting process and hence not observed in serum. We have developed, characterised and validated an efficient, highly sensitive and specific method for the quantification of S1P in biological material.


A novel plant-made monoclonal antibody enhances the synergetic potency of an antibody cocktail against the SARS-CoV-2 Omicron variant.
This study describes a novel, neutralizing monoclonal antibody (mAb), 11D7, discovered by mouse immunization and hybridoma generation, against the parental Wuhan-Hu-1 RBD of SARS-CoV-2. We further developed this mAb into a chimeric human IgG and recombinantly expressed it in plants to produce a mAb with human-like, highly homogenous N-linked glycans that has potential to impart greater potency and safety as a therapeutic. The epitope of 11D7 was mapped by competitive binding with well-characterized mAbs, suggesting that it is a Class 4 RBD-binding mAb that binds to the RBD outside the ACE2 binding site. Of note, 11D7 maintains recognition against the B.1.1.529 (Omicron) RBD, as well neutralizing activity. We also provide evidence that this novel mAb may be useful in providing additional synergy to established antibody cocktails, such as Evusheld™ containing the antibodies tixagevimab and cilgavimab, against the Omicron variant. Taken together, 11D7 is a unique mAb that neutralizes SARS-CoV-2 through a mechanism that is not typical among developed therapeutic mAbs and by being produced in ΔXFT Nicotiana benthamiana plants, highlights the potential of plants to be an economic and safety-friendly alternative platform for generating mAbs to address the evolving SARS-CoV-2 crisis.


Custom implants for medialization laryngoplasty: a model that considers tissue compression.
Unilateral vocal fold paralysis can be treated with surgical medialization with a carved silastic implant. One challenge to this approach is anticipating the dimensions of the implant for adequate medialization. The purpose of this study was to develop a predictive model for implant design based on a patient's unique laryngeal anatomy and considering compression of the vocal fold. Retrospective chart review, prospective cadaver study, and prospective patient study. A retrospective chart review was performed on patients who received silastic medialization laryngoplasty with favorable outcome and who had preoperative computed tomography. Data including Voice Handicap Index, maximum phonation time, and implant dimensions were collected from medical records, and laryngeal measurements were taken from preoperative imaging. Measurements were taken from computed tomography scans of three cadavers who underwent laryngoplasty for this study. Tissue compression (TC) was calculated and analyzed. A model to predict successful implant dimensions was developed and applied prospectively in 16 patients. Eleven patients from the chart review and three cadavers were included. Of all laryngeal metrics, width of the vocal fold at maximal medialization was most strongly correlated to TC (r = 0.728). Linear regression was performed (y = 0.50x - 1.2, R(2) = 0.53, P = 0.005, F = 12.39). Of the prospective patients, 15 of 16 demonstrated complete glottis closure with the premeasured silastic implant. Vocal fold compression by silastic implants is linearly correlated with vocal fold-width at maximal medialization. A predictive formula was generated to anticipate TC and was successful in designing custom implants for patients.


Wireless intra-brain communication for image transmission through mouse brain.
We demonstrate wireless image data transmission through a mouse brain. The transmission characteristics of mouse brain is measured. By inserting electrodes into the brain, the transmission efficiency is drastically increased. An AM signal modulated with the image data from an implantable image sensor was launched into the brain and the received signal was demodulated. The data was successfully transmitted through the brain and the image was reproduced.


Lipid nanoparticle technologies for the study of G protein-coupled receptors in lipid environments.
G protein-coupled receptors (GPCRs) are a large family of integral membrane proteins which conduct a wide range of biological roles and represent significant drug targets. Most biophysical and structural studies of GPCRs have been conducted on detergent-solubilised receptors, and it is clear that detergents can have detrimental effects on GPCR function. Simultaneously, there is increasing appreciation of roles for specific lipids in modulation of GPCR function. Lipid nanoparticles such as nanodiscs and styrene maleic acid lipid particles (SMALPs) offer opportunities to study integral membrane proteins in lipid environments, in a form that is soluble and amenable to structural and biophysical experiments. Here, we review the application of lipid nanoparticle technologies to the study of GPCRs, assessing the relative merits and limitations of each system. We highlight how these technologies can provide superior platforms to detergents for structural and biophysical studies of GPCRs and inform on roles for protein-lipid interactions in GPCR function.


Dissecting the phenotypic components and genetic architecture of maize stem vascular bundles using high-throughput phenotypic analysis.
High-throughput phenotyping is increasingly becoming an important tool for rapid advancement of genetic gain in breeding programmes. Manual phenotyping of vascular bundles is tedious and time-consuming, which lags behind the rapid development of functional genomics in maize. More robust and automated techniques of phenotyping vascular bundles traits at high-throughput are urgently needed for large crop populations. In this study, we developed a standard process for stem micro-CT data acquisition and an automatic CT image process pipeline to obtain vascular bundle traits of stems including geometry-related, morphology-related and distribution-related traits. Next, we analysed the phenotypic variation of stem vascular bundles between natural population subgroup (480 inbred lines) based on 48 comprehensively phenotypic information. Also, the first database for stem micro-phenotypes, MaizeSPD, was established, storing 554 pieces of basic information of maize inbred lines, 523 pieces of experimental information, 1008 pieces of CT scanning images and processed images, and 24 192 pieces of phenotypic data. Combined with genome-wide association studies (GWASs), a total of 1562 significant single nucleotide polymorphism (SNPs) were identified for 30 stem micro-phenotypic traits, and 84 unique genes of 20 traits such as VBNum, VBAvArea and PZVBDensity were detected. Candidate genes identified by GWAS mainly encode enzymes involved in cell wall metabolism, transcription factors, protein kinase and protein related to plant signal transduction and stress response. The results presented here will advance our knowledge about phenotypic trait components of stem vascular bundles and provide useful information for understanding the genetic controls of vascular bundle formation and development.


[Study of the method to measure the diameter of retinal vessels].
To estimate the objective and repeatable method for measurements of the retinal vessels' diameter. The direct reason for creating this method was to use it to evaluate the influence of exogenous estrogens on the retinal blood vessels. Based on modern computer systems, using funduscamera Canon NF 505, the model of active matrix for repeatable method for estimating the changes in vessels' diameter, was elaborated. After statistical analysis of a material of 3000 measurements, the degree of objectiveness and recurrence and the potential scale for errors was acknowledged. This method enables the estimation of the changes in retinal vessels diameter caused by many different factors in comparison to initial observations (set as 100%) in every analysed case. Presented method can be successfully used for objective, reliable evaluation of changes in the blood vessels diameter caused by many different factors.


Heart failure with preserved ejection fraction.
Half of all heart failure patients have preserved ejection fraction, but there is no established therapy for this patient group. Effective heart failure therapy depends on an understanding of the underlying pathophysiology. This article presents an updated review of knowledge on the causal mechanisms underlying heart failure with preserved ejection fraction (HFpEF). Articles were found by means of a literature search in PubMed. The search combination "heart failure with preserved ejection fraction" OR "HFpEF" OR "diastolic heart failure") AND ("mechanisms" OR "hypertrophy" OR "inflammation") yielded 603 hits on 6 April 2017. Relevant articles on causal mechanisms were read in full text. In recent years there has been a paradigm shift with respect to understanding of the pathophysiology of HFpEF. Concentric hypertrophy of the left ventricle with subsequent diastolic dysfunction had long been recognised as an important disease mechanism, but recent research has identified other factors that also contribute to the condition. These include systolic dysfunction, abnormal regulation of heart rhythm, pathological vascular stiffness, autonomic dysfunction and peripheral vasculopathy. Several studies have suggested that comorbidity plays a part by inducing a systemic proinflammatory response which results in multi-organ dysfunction. The pathophysiological picture of HFpEF indicates that the condition resembles a syndrome more than an isolated cardiac disorder. A stronger focus on comorbidity may lead to new diagnostic and therapeutic options.


Copper and cadmium impair sperm performance, fertilization and hatching of oocytes from Amazonian fish Colossoma macropomum.
The contamination of aquatic environments by transition metals can have a direct influence on the reproductive process of several organisms in the aquatic biota. This study aimed to evaluate the effect of cadmium and copper on the sperm of tambaqui (Colossoma macropomum). Male (n = 4) and female (n = 4) specimens of C. macropomum were induced to spermiation and ovulation, with sperm being activated in the following media: 0; 0.6; 1.2 and 1.8 mg/L of cadmium (CdCl2) and 0; 0.4; 0.8 and 1.2 mg/L of copper (CuCl2). Sperm quality was assessed through time (s) and motility rate (%), superoxide dismutase (SOD) and glutathione S-transferase (GST) activities, lipoperoxidation levels (LPO), and morphological characteristics. In parallel, the effects of these metals on the rate of fertilization and hatching of the oocytes were evaluated. The duration and motility rate of sperm were longer in the control treatment, 85.67 ± 11.01 s; 90 ± 0.01%, and progressively decreased to 44.67 ± 4.16 s and 60 ± 5%, respectively, in concentrations of 1.8 mg/L (44.67 ± 4.16 s; 60 ± 5%) of CdCl2 and to 65.67 ± 3.30 s; 70 ± 5%, respectively, in concentrations of 0.8 mg/L of CuCl2. We observed an increase in the activity of the SOD enzyme in sperm cells exposed to 1.2 mg/L of CdCl2. The LPO levels were increased significantly in sperm cells exposed to 1.2 and 1.8 mg/L of CdCl2 and 0.8 mg/L of CuCl2. Fertilization and hatching were severely impaired in the presence of Cd and Cu. These data indicate that environments contaminated with cadmium and copper harm the gametes of C. macropomum.


Disseminated lupus vulgaris presenting as granulomatous folliculitis.
A 69-year-old man presented with a 3-year history of scattered, grouped, asymptomatic follicular papules, pustules, and nodules tending toward coalescence into large geographic aggregates. Repeated cutaneous biopsy specimens showed granulomatous folliculitis with negative Ziehl-Neelsen stains. Finally, biopsy material submitted for mycobacterial culture grew Mycobacterium tuberculosis. No evidence of active systemic tuberculosis was found. The patient had a very rare form of tuberculosis, disseminated lupus vulgaris, presenting with granulomatous folliculitis, which is usually not seen in lupus vulgaris. The lesions resolved after an 18-month course of isoniazid and rifampin. The unusual clinical and histologic presentation as well as occasional partial remissions following a variety of nonspecific empiric therapies delayed diagnosis despite multiple evaluations. This case illustrates the importance of obtaining mycobacterial cultures from skin biopsy specimens in addition to special stains whenever cutaneous tuberculosis is suspected.


Interleukin-1 beta induction of neuron apoptosis depends on p38 mitogen-activated protein kinase activity after spinal cord injury.
Interleukin-1 beta (IL-1beta) has been implicated as an extracellular signal in the initiation of apoptosis in neurons and oligodendrocytes after spinal cord injury (SCI). To further characterize the apoptotic cascade initiated by IL-1beta after SCI, we examined the expression of IL-1beta, p38 mitogen-activated protein kinase (p38 MAPK) and caspase-3 after SCI, and further investigated whether p38 MAPK was involved in neuron apoptosis induced by IL-1beta. Adult rats were given contusion SCI at the T-10 vertebrae level with a weight-drop impactor (10 g weight dropped 25.0 mm). The expression levels of IL-1beta, p38 MAPK and caspase-3 after SCI were assessed with Western blots, immunohistochemistry staining, and real time reverse transcription polymerase chain reactions (RT-PCR). Neuron apoptosis was assessed with the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) method. Increased levels of IL-1beta and p38 MAPK were observed soon after injury, with a peak in expression levels within 6 h of injury. By 24 h after injury, caspase-3 expression was markedly increased in the injured spinal cord. TUNEL-positive cells were first observed in the lesioned area 6 h after SCI. The largest number of TUNEL-positive cells was observed at 24 h post-SCI. Intrathecal injection of the IL-1 receptor antagonist IL-1Ra significantly reduced expression of p38 MAPK and caspase-3, and reduced the number of TUNEL-positive cells. Moreover, intrathecal injection of an inhibitor of p38 MAPK, SB203580, also significantly reduced the expression of caspase-3, and reduced the number of TUNEL-positive cells in the injured spinal cord. The p38MAPK signaling pathway plays an important role in IL-1beta mediated induction of neuron apoptosis following SCI in rats.


Intensive care unit nurses' perceptions of debriefing after critical incidents: A qualitative descriptive study.
Intensive care unit (ICU) nurses are exposed to critical incidents daily at their workplace, which may have long-term physical and psychological impacts. Despite the growing evidence supporting clinical debriefing in health care to prevent these impacts, a scarcity of literature exists to support its use in the adult intensive care setting. The objective of this study was to explore nurses' perceptions of clinical debriefing after critical incidents in an adult ICU. A qualitative descriptive design was utilised. Thematic analysis of data from individual semistructured interviews with six ICU nurses was undertaken. In this study, two themes were identified. Firstly, participants valued hot debriefing after critical incidents for the key reasons of having an opportunity to reflect on and learn from a critical incident and reduce normalisation of stressful situations. Secondly, when logistical factors such as communication, timing, and location were not considered, the attendance at debriefings was negatively influenced. Participants identified that ICU nurses commonly prioritised patient tasks over attending a debrief; therefore, teamwork and flexibility with logistics was crucial. Hot debriefing, of a short duration and close to the time of the event, was valued and played an important role in staff wellbeing and self-care, contributing to preventing self-blame and normalisation of stressful situations. A clearer definition of the term along with greater recognition of types of events that could be considered critical incidents is required for staff support after critical incidents in the complex intensive care setting.


The maximum chemiluminescence intensity predicts severe neutropenia in gemcitabine-treated patients with pancreatic or biliary tract cancer.
To assess the predictive ability of the maximum chemiluminescence intensity (CImax) for severe neutropenia (SN) during neoadjuvant chemo(radio)therapy [NAC(RT)] in patients with advanced pancreatic or biliary tract cancer. Clinicopathological variables and blood test data before NAC(RT) were evaluated in 64 patients with advanced pancreatic or biliary tract cancer who received gemcitabine plus tegafur/gimeracil/oteracil as NAC(RT). Thirty-nine patients (60.9%) developed Grade 3-4 SN. The median time between commencing NAC(RT) and the onset of SN was 15 (range 10-36) days. SN occurred during the NAC period, not the RT period. The CImax, neutrophil count, serum interleukin-6 level, C-reactive protein level, complement C3 titer, serum complement titer, and 50.0% hemolytic unit of complement before NAC(RT) were significantly lower in patients with SN than in those without SN (P < 0.05). Multivariate analysis confirmed the CImax to be the sole independent predictor of SN (P < 0.05). The optimal threshold for the CImax was 46,000 RLU/s. The sensitivity and specificity were 46.2% and 80.0%, respectively. Majority of the patients (81.8%) with a low CImax before NAC(RT) experienced SN during NAC(RT). CImax before NAC(RT) predicts SN during NAC(RT) in patients with advanced pancreatic or biliary tract cancer.


5alpha-reduced neuroactive steroids alleviate thermal and mechanical hyperalgesia in rats with neuropathic pain.
5alpha-reduced neuroactive steroids with selective modulatory action in vitro on T or combined modulatory action on T and GABA(A) currents present in peripheral sensory neurons have been shown to induce potent peripheral analgesia in vivo in intact animals. Although the role of T and GABA(A) currents in pathophysiology of neuropathic pain (NPP) is not established, it appears that blockade of T currents and/or potentiation of GABA(A) currents could be beneficial in the management of NPP. To study the potential usefulness of 5alpha-reduced neuroactive steroids in alleviating NPP, we selected two newly synthesized steroids-ECN and CDNC24-with a selective blocking effect on T currents and a selective potentiating effect on GABA(A) currents, respectively, and commercial analogs-alphaxalone and 3alpha5alphaP-with the effects on both ion channels. We used a sciatic nerve ligation model to induce thermal and mechanical hyperalgesia in adult rats and tested peripheral thermal and mechanical nociception following local injection of neuroactive steroids into the peripheral receptive fields of a ligated hind paw. We found that 5alpha-reduced neuroactive steroids alleviate thermal and mechanical hyperalgesia in NPP rats. ECN and CDNC24 were more selective in alleviating thermal nociception in NPP than in sham animals when compared to 3alpha5alphaP and alphaxalone although the anti-nociceptive effect induced by 3alpha5alphaP and alphaxalone was more profound. CDNC24 was most selective since it had very minimal anti-nociceptive effect in sham animals but a very profound anti-nociceptive effect in NPP animals suggesting that, under pathological conditions, peripheral GABA(A) receptors might be an attractive therapeutic target.


[A preliminary study of pentanucleotide repeat polymorphism of apolipoprotein(a) gene in coronary atherosclerotic heart disease].
This study inquired into the relationship between a pentanucleotide repeats(PNR) polymorphism of the apolipoprotein(a)[apo(a)] gene and coronary atherosclerotic heart disease(CHD) in Chinese Han nationality. PNR polymorphism of the apo(a) gene from 165 cases of CHD and 153 normal individuals were analyzed by polymerase chain reaction(PCR)-denature polyacrylamide gel electrophoresis-silver stain. The frequencies of (TTTTA)(5/8) genotype (0.188) and (TTTTA)(5) allele (0.115) in CHD group were remarkably higher than those in control group (0.039, 0.026)(P<0.01, P<0.05). These findings indicate that the PNR polymorphism of apo(a) is associated with the susceptibility to CHD, which may be involved in the development of CHD.


Analysis of interactions between cinnamycin and biomimetic membranes.
The interaction between the cinnamycin and the biomimetic membranes was studied using the atomic force microscope(AFM). The bilayer was composed of the monolayer tethered on the gold surface and the outer layer fused with the vesicles on the monolayer. The vesicles were prepared at the desired ratio of dioleoylphosphatidylethanolamine(DOPE) to dioleoylphosphatidylcholine(DOPC). On the bilayer, the surface force measurement was performed with the cinnamycin immobilized covalently on the tip surface. The immobilization led to the presence of the adhesion, which was found while the tip was retracted from the bilayer. In addition, the magnitude of the adhesive force was changed with respect to the composition of DOPE in the outer layer. The difference in the adhesion may be attributed to the mean-molecular-area of DOPE and the specific-binding density on the outer layer. Furthermore, the analysis of the rupture force with respect to the loading rate indicated that the rupture length was around 0.1∼0.13 nm, which was similar to that of a van der Waals bond.


De novo partial 2q3 trisomy/distal 7p22 monosomy in a malformed newborn with 7p deletion phenotype and craniosynostosis.
In the present paper a malformed male newborn is presented with de novo 2q3 trisomy/distal 7p22 monosomy and typical clinical findings of 7p deletion syndrome including trigonocephaly.


Low-dose ketamine infusions reduce opioid use in pediatric and young adult oncology patients.
Ketamine is an NMDA-receptor antagonist with analgesic and opioid-sparing properties. Although well studied in adults, more robust evidence supporting ketamine's use for pediatric pain management is needed. This retrospective study evaluates ketamine's opioid-sparing effectiveness in pediatric and young adult oncology and hematology patients. Continuous ketamine infusions administered for pain management between 2010-2020 were reviewed. Data including demographic characteristics, oncology/hematology and pain diagnoses, concurrent pain medications, and ketamine infusions' dose and duration were collected. Opioid consumption data based on delivery via patient-controlled analgesia were collected 1 day before (D1), all days during (cumulatively named D2), and 1 day after (D3) ketamine infusions and calculated as morphine-equivalent doses (mg/kg/day). Data were reported for the entire study group as well as for distinct oncology and end-of-life categories, and short-term acute pain circumstances which included vaso-occlusive crises in hematology patients. Side effects were reviewed. Significantly lower daily opioid consumption was noted in the oncology group, while decreases were not significant in the end-of-life group and in the overall study population. The acute pain group did not show an opioid reduction associated with the ketamine infusions. A largely tolerable side-effect profile was observed, with no differences among each group's incidence. Ketamine infusions were associated with significantly reduced opioid consumption for oncology patients. The opioid-sparing effects of ketamine may vary according to clinical diagnoses and circumstances of use. Overall, low-dose ketamine infusions present an acceptable safety profile in pediatric and young adult patients; nevertheless, individual risks and benefits should be considered.


Correction: Predictors of Playing Augmented Reality Mobile Games While Walking Based on the Theory of Planned Behavior: Web-Based Survey.
[This corrects the article DOI: 10.2196/mhealth.8470.].


A structural basis for the amphiphilic character of alginates - Implications for membrane fouling.
Ostensibly hydrophilic alginates are known to foul hydrophobic membranes, under various conditions. Here, controlled experiments have been conducted at high and low pH on the fouling of a polypropylene membrane by alginate and the results suggest that the observed fouling is due to an intrinsic property of the alginate. Thus quantum chemical calculations on the M and G monomers of alginate reveal that M adopts an equilibrium geometry that is hydrophilic on one face and hydrophobic on the other, i.e. is potentially amphiphilic. Molecular dynamics simulations on short alginate chains of different sequences interacting with a modelled polypropylene surface, show that this characteristic is carried over to the polymer and results in hydrophobic patches along the chain that facilitate attractive interactions with the polypropylene surface. This concept is buttressed by an analysis of the binding characteristics of a previously reported X-ray structure of the mannuronan C-5 epimerase AlgE4 enzyme.


Pharmacological characterization of AS2690168, a novel small molecule RANKL signal transduction inhibitor.
Pathological osteolysis is associated with excessive bone resorption by activated osteoclasts. Given that receptor activator of NF-kB and its ligand (RANKL) are key players in the differentiation and activation of osteoclasts, the RANKL/RANK signaling pathway is considered a promising target for the development of effective osteoclastogenesis inhibitors. We previously found that the orally available compound, AS2690168, suppresses RANKL-induced osteoclastogenesis of RAW264 cells. In this report, we further characterized the pharmacological profiles of AS2690168 in vitro and in vivo. AS2690168 suppressed soluble RANKL (sRANKL)-induced NFATc1 mRNA expression in RAW264 cells at 0.3 and 3.0 μM. It also suppressed calcium release from parathyroid hormone-stimulated mouse calvaria with an IC50 value of 0.46 μM. Oral administration of AS2690168 completely suppressed the decrease in femoral bone mineral content in an sRANKL-induced osteopenic mice model at 3.0 mg/kg. It also significantly suppressed the decrease in femoral bone mineral density and increase in serum tartrate-resistant acid phosphatase-5b levels in ovariectomized rats at doses of 0.3, 1 and 3 mg/kg. Finally, AS260168 suppressed the increase in urine deoxypyridinoline in a rat prednisolone-induced osteoporosis model at 10 mg/kg. These results suggest that AS2690168 is a promising treatment for bone disorders with excessive bone resorption.


Acupuncture benefits to women with recurrent implantation failure: A propensity score-matched cohort study.
The current study aims to assess the effectiveness of acupuncture in improving the live birth rate (LBR), ongoing pregnancy rate (OPR), clinical pregnancy rate (CPR), biochemical pregnancy rate (BPR), and pregnancy loss (early abortion rate, late abortion rate, ectopic pregnancy rate) in patients with recurrent implantation failure (RIF). This retrospective study compares the outcomes of patients with RIF who underwent frozen embryo transfer (FET) with or without acupuncture. The medical records of patients diagnosed with RIF and visiting Chengdu Xi'nan Gynecological Hospital between January 2018 and June 2021 were reviewed. The Chengdu Xi'nan Gynecological Hospital Ethics Committee approved this retrospective study (No. 2021-029). A total of 923 patients with RIF who underwent FET were included in this study. The patients were divided into two groups: the Acupuncture (n = 303) and the Non-acupuncture groups (n = 620). The Acupuncture group consisted of 303 RIF patients who received acupuncture therapy in addition to standard hormone replacement therapy (HRT)/delayed hormone replacement therapy (d-HRT) for FET. The Non-acupuncture group consisted of 620 RIF patients who received only standard HRT/d-HRT for FET. The primary outcome was the LBR. The secondary outcome referred to OPR, CPR, BPR, and pregnancy loss. The Acupuncture group had significantly higher BPR (P = 0.08) and CPR (P = 0.049) than the Non-acupuncture group. A potentially higher LBR (P = 0.16) and OPR (P = 0.248) were observed in the Acupuncture group than in the Non-acupuncture group. However, the survival analysis did not show that acupuncture significantly promoted live birth. Acupuncture is an appropriate adjunctive technique in the in vitro fertilization process as it improves biochemical and clinical pregnancies. Therefore, it is necessary to be cautious about the role of acupuncture throughout the whole pregnancy cycle.


Genesis of "negative pressure" during hanging drop; the answer is positive pressure.
A drop of saline placed within the hub of an epidural needle is sucked in when the needle is advanced and penetrates through a polyurethane foam cube. This phenomenon might be explained by the release of compressed air inside of the needle into the air. The positive pressure inside of the needle may be generated by advancing of the needle. To prove this hypothesis, the pressure inside of the needle was measured during needle advancement, and it increased gradually and then dropped to zero suddenly when the needle fully penetrated the polyurethane foam block. We can speculate that the same phenomenon occurs during hanging-drop method of epidural anesthesia, and this may occur regardless of whether epidural subatmospheric pressure exists or not.


Actions of thiazide on vitamin D metabolism: a controlled therapeutic trial in normal women early in the postmenopause.
The effect of thiazide on vitamin D metabolism in normal postmenopausal women was studied during a twelve-month placebo-controlled clinical study. Nineteen healthy women in their early menopause were randomized for treatment with bendroflumethiazide (5 mg/d) (n = 11) or placebo (n = 8) for twelve months. All participants were given a calcium supplement of 0.5 g/d throughout the study. A significant increase (p less than 0.01) in the serum concentration of 24,25-dihydroxycholecalciferol was observed in the thiazide group. Moreover, this group showed a tendency toward decreased serum 1,25-dihydroxycholecalciferol, whereas the mean serum 25-hydroxycholecalciferol was unchanged. Except for a highly significant decrease in urinary calcium in the thiazide group (P less than 0.01) all other biochemical indices of calcium metabolism were unchanged. The present data indicate that thiazide given to early postmenopausal women has a primary effect on the renal tubules followed by a secondary change in vitamin D metabolism leading to an increase in serum 24,25-dihydroxycholecalciferol.


Proteoglycan Endocan: A multifaceted therapeutic target in Cancer.
Endocan is known to be a circulating dermatan sulfate proteoglycan that regulates endothelial cell function. Dysregulation of endocan expression is observed not only in the tumor vasculature but also in cancer cells. Accumulating evidence has revealed that disordered endocan facilitates cancer progression via enhancing cancer cell proliferation, cell mobility, and cancer stemness properties. Recently, various interacting proteins and diverse subcellular localizations of endocan were identified in cancer cells. Herein, we summarize the application of endocan in cancer diagnoses and prognoses using serum and tumor specimens. We further discuss that the aberrant molecular characteristics of endocan may be due to the mislocalization of endocan in cancer cells. Defining the specific cellular roles of endocan will provide a promising diagnostic factor and therapeutic target for cancer patients.


Validity of the Brazilian version of the COPD assessment test in patients with chronic obstructive pulmonary disease.
COPD Assessment Test (CAT) is a questionnaire defined as one of the main options in the assessment of the impact of Chronic Obstructive Pulmonary Disease on patients' life, according to the Global Initiative on Obstructive Lung Diseases. However, the Consensus-based Standards for the Selection of Health Measurement Instruments-COSMIN 2018 has recently determined a correlation coefficient greater than 0.70 and an optimal sample size must have a hundred subjects, hence, validity of the Brazilian version of this instrument is still not yet proven. To verify the validity of the Brazilian version of the Questionnaire CAT in patients with Chronic Obstructive Pulmonary Disease, comparing it to the Saint George's Respiratory Questionnaire (SGRQ) and correlate it with other outcomes. Observational transversal study with 101 COPD diagnosed subjects who underwent anthropometric data collection, life habits, CAT and SGRQ questionnaires, modified Medical Research Council (mMRC) dyspnoea scale. Spirometry, six-minute walking test (6MWT) and body mass index, Dyspnoea and exercise capacity (BODE). Each evaluative measure was compared to the total CAT score. The validity was considered for a significant Pearson correlation coefficient (r) greater than 0.7. It was observed a strong correlation between the total CAT score and the total SGRQ score (r = 0.703; P < 0.001), as well as moderate correlation with mMRC (r = 0.518; P < 0.001). SGRQ Symptoms (r = 0.627; P < 0.001). SGRQ Activity (r = 0.636; P < 0.001) and SGRQ Impact (r = 0.655; P < 0.001). There were similar correlations between CAT and other variables (6MWT, mMRC and BODE) and SGRQ and the same variables. The CAT questionnaire is a valid, reliable and easy-to-use instrument for assessing symptoms and can record the impact of COPD on the patient's quality of life.


Differential characterization of hepatic tumors in MR imaging by burst-released Mn2+-ions from hollow manganese-silicate nanoparticles in the liver.
Gd3+-based contrast agents monopolize in the clinical MR imaging-based diagnosis of hepatic tumors, however, the inherent toxicity by the released Gd3+-ions triggered an urgent demand for safer alternatives. Here, we demonstrate hollow manganese silicate nanoparticles (HMS), which exert burst-release of Mn2+-ions by switching to physiological acidic condition, exhibiting high effectiveness in hepatic tumor characterization as liver-specific MR contrast agent through the in-depth in vivo MR imaging study and immunohistochemical investigations with three hepatic tumor models (e.g., hepatocellular carcinoma, neuroendocrine carcinoma, adenocarcinoma). Their characteristic time-sequential enhancement patterns in HMS-enhanced MR imaging with improved conspicuity reflect their biological features such as vascularity, cellularity, mitochondrial activity and hepatocellular specificity, and thus allow the disease-specific characterization of various hepatic tumors. HMS-enhanced MR imaging with necrotic hepatocellular carcinoma model suggested the good correlation of the extent of tumor necrosis with residual mitochondrial activity. Such multi-responsive spatio-biological distribution and function of HMS resulting in time-dependent bioimaging coupled with low systemic toxicity sets the clinical potential to accurate diagnosis and therapeutic response in various hepatic tumors.


Fat-plug and paper-patch myringoplasty in rats.
The purpose of this study is to investigate the recovery rate in paper-patch and fat-plug myringoplasty in rats. Small and large perforations were inflicted on the tympanic membranes of 60 rats. Fat-plug and paper-patch myringoplasties were performed to different groups and another group was left for spontaneous healing. We found that for small perforations, the recovery rate was 94.7% in fat-plug myringoplasty, 94.4% in paper-patch myringoplasty, and 66.6% in control group. The recovery rates in large perforations were 52.9%, 56.2%, and 26.6%, respectively. On the other hand, in larger perforations, paper-patch or fat-plug myringoplasty have not been found effective. We believe that due to significant operational advantages, fat-plug or paper-patch myringoplasty can be suggested for the reconstruction of small and dry perforations of the tympanic membrane.


Metformin as a promising target for DPP4 expression: computational modeling and experimental validation.
Metformin is a regularly prescribed and low-cost generic medication. Metformin has been proposed as a target for Dipeptidyl-peptidase 4 (DPP4) expression in various clinical disorders. We provide insilco investigations on molecular docking and dynamic modeling of metformin and DPP4 potential interactions. Moreover, we conducted bioinformatic studies to highlight the clinical significance of DPP4 expression and mutation in various types of malignancies, as well as the invasion of different immune cells into the tumor microenvironment. We believe the present proposal's findings have crucial implications for understanding how metformin may confer health advantages by targeting DPP4 expression in malignancies.


238Pu and 239+240Pu inventory and distribution through the lower Rhone valley terrestrial environment (Southern France).
Because of atmospheric global fallout due to thermonuclear tests performed between 1945 and 1980 and to the American SNAP 9A satellite explosion in 1964, plutonium's long-lived alpha emitter isotopes ((238)Pu, (239)Pu and (240)Pu) can be found everywhere in the environment. In the soils of the lower Rhone valley, over a surface area of approximately 11,000 km(2), existing results allow estimation of the total inventory resulting from this origin of 551 GBq as regards these radionuclides. The Marcoule Nuclear Reprocessing Plant (NRP) has been releasing (238)Pu and (239+240)Pu into the environment for over 40 years in the region and is as such a second likely source for Pu input. This article gives a global review of plutonium distribution in this particular region. It is shown that releases from Marcoule, although accounting for less than 2% of the total Pu inputs into the terrestrial environment, are responsible for localised and measurable Pu enrichments close to the facility (+2.8 GBq), and in the Rhone delta where sediments from the river settled during the past floods (+0.1 GBq). Irrigating with water from the Rhone River also allowed the transfer of approximately 3.8 GBq of (238)Pu and (239+240)Pu to the soils for the whole 1960-1998 period. These inputs, distributed over wide irrigated surface areas, do not induce significant increases of soil Pu-activity levels. In a second section, this study confirms that most of the plutonium existing in the terrestrial environment is accumulated in the soil. Less than 0.1% of the activities exist currently in the plant compartments, and the flows exported by agricultural productions are very low, although we are interested here in a French region where agriculture prevails.


Examining domains of community health nurse satisfaction and motivation: results from a mixed-methods baseline evaluation in rural Ghana.
A strong health system requires a competent and caring workforce. A more satisfied and motivated health workforce should be more willing to serve in difficult areas, have lower turnover, and theoretically provide better care to patients. This paper examines the motivation, satisfaction, and correlation with clinical knowledge, of community health nurses (CHNs), a cadre of provider focused on maternal, newborn and child health in rural Ghana. This study employed three methods of evaluation. Two quantitative measurements were used: (1) a survey of health worker satisfaction and motivation and (2) a clinical knowledge assessment focusing on maternal, newborn and child health. Both were administered to all rostered CHNs working in the five sampled districts in the Greater Accra and Volta regions in Eastern Ghana (N = 205). Qualitative interviews (N = 29) and focus group discussions (N = 4) were held with selected CHNs in the same districts. These data were analysed using NVivo (Version 10) and Stata (Version 13.0) based on domains of extrinsic and intrinsic motivation including general satisfaction, work environment and access to resources, respect and recognition received and opportunities for advancement. CHNs desired more training, especially those who were posted at the community level (a Community-based Health Planning and Services post or "CHPS") versus at a health facility. CHNs working at CHPS believed their work to be more difficult than those posted at health facilities, due to challenges associated with foot travel to visit patients at home, and they were more likely to report having insufficient resources to do their jobs (48% vs 36%). However, CHNs posted at health facilities were more likely to report insufficient opportunities for career advancement than the CHPS nurses (49% vs 33%). CHNs generally reported good relationships with colleagues and being respected by patients but desired more respect from supervisors. The median score on the knowledge assessment was 78%. On average, subgroups of CHNs with different reported levels of satisfaction did not perform differently on the knowledge assessment. CHNs in Ghana were satisfied overall but desired more training, more guidance and supervision, fair pay and opportunities to advance in their career. Improving health worker satisfaction and morale may be important for health worker retention and certain aspects of care but may not have a significant influence on clinical knowledge or performance.


Inhibition of Arabidopsis stomatal development by plastoquinone oxidation.
Stomata are the pores in the epidermal surface of plant leaves that regulate the exchange of water and CO2 with the environment thus controlling leaf gas exchange.1 In the model dicot plant Arabidopsis thaliana, the transcription factors SPEECHLESS (SPCH) and MUTE sequentially control formative divisions in the stomatal lineage by forming heterodimers with ICE1.2 SPCH regulates entry into the stomatal lineage and its stability or activity is regulated by a mitogen-activated protein kinase (MAPK) signaling cascade, mediated by its interaction with ICE1.3-6 This MAPK pathway is regulated by extracellular epidermal patterning factor (EPFs) peptides, which bind a transmembrane receptor complex to inhibit (EPF1 and EPF2) or promote (STOMAGEN/EPFL9) stomatal development.7-9 MUTE controls the transition to guard mother cell identity and is regulated by the HD-ZIP transcription factor HDG2, which is expressed exclusively in stomatal lineage cells.10,11 Light signals acting through phytochrome and cryptochrome photoreceptors positively regulate stomatal development in response to increased irradiance.12,13 Here we report that stomatal development is also regulated by the redox state of the photosynthetic electron transport chain (PETC). Oxidation of the plastoquinone (PQ) pool inhibits stomatal development by negatively regulating SPCH and MUTE expression. This mechanism is dependent on MPK6 and forms part of the response to lowering irradiance, which is distinct to the photoreceptor dependent response to increasing irradiance. Our results show that environmental signals can act through the PETC, demonstrating that photosynthetic signals regulate the development of the pores through which CO2 enters the leaf.


Does right thoracotomy increase the risk of mitral valve reoperation?
The study objective was to determine whether a right thoracotomy approach increases the risk of mitral valve reoperation. Between January of 1993 and January of 2004, 2469 patients with mitral valve disease underwent 2570 reoperations (1508 replacements, 1062 repairs). The approach was median sternotomy in 2444 patients, right thoracotomy in 80 patients, and other in 46 patients. Multivariable logistic regression was used to identify factors associated with median sternotomy versus right thoracotomy, mitral valve repair versus replacement, hospital death, and stroke. Factors favoring median sternotomy (P < .03) included coronary artery bypass grafting (30% vs 2%), aortic valve replacement (39% vs 2%), tricuspid valve repair (27% vs 13%), fewer previous cardiac operations, more recent reoperation, and no prior left internal thoracic artery graft. These factors were used to construct a propensity score for risk-adjusting outcomes. Hospital mortality was 6.7% (163/2444) for the median sternotomy approach and 6.3% (5/80) for the thoracotomy approach (P = .9). Risk factors (P < .04) included earlier surgery date, higher New York Heart Association class, emergency operation, multiple reoperations, and mitral valve replacement. Stroke occurred in 66 patients (2.7%) who underwent a median sternotomy and in 6 patients (7.5%) who underwent a thoracotomy (P = .006). Mitral valve replacement (vs repair) was more common in those receiving a thoracotomy (P < .04). Compared with median sternotomy, right thoracotomy is associated with a higher occurrence of stroke and less frequent mitral valve repair. Specific strategies for conducting the operation should be used to reduce the risk of stroke when right thoracotomy is used for mitral valve reoperation. In most instances, repeat median sternotomy, with its better exposure and greater latitude for concomitant procedures, is preferred.


Cathepsin K inhibitors prevent matrix-derived growth factor degradation by human osteoclasts.
The coupling between bone formation and resorption creates a therapeutic impasse in osteoporosis: antiresorptive therapy halts bone loss, but also inhibits bone formation, and therefore does not cure the condition. Surprisingly, recent preliminary reports suggest that inhibition of resorption by cathepsin K (CathK) inhibitors augments bone formation. Uniquely amongst resorption-inhibitors, CathK-inhibitors suppress degradation of the organic matrix of bone while allowing demineralization. We hypothesized that these unique characteristics might explain a capacity of CathK inhibitors to enhance bone formation: the inhibitors might prevent degradation not only of collagen, but also other proteins, including growth factors embedded in matrix. We tested this hypothesis using osteocalcin and insulin-like growth factor I (IGF-I) as examples of matrix-embedded proteins, and found that CathK-inhibitors, unlike other resorption-inhibitors, dramatically increased the concentrations of these matrix-derived proteins in supernatants of osteoclasts on bone, most likely through protection against intracellular degradation. We found that protons are both necessary and sufficient for the release of IGF-I from bone matrix, and that recombinant CathK can degrade both marker proteins. In the presence of a CathK-inhibitor, the amount of IGF-I released from matrix substantially exceeded the amount secreted by osteoclasts. CathK-inhibition similarly augmented bone morphogenetic protein (BMP)-2 release. Lastly, MC3T3-E1 numbers were greater after co-culture with osteoclasts on bone with versus without CathK-inhibitor, showing that, in the presence of CathK-inhibitor, osteoclasts release biologically-significant quantities of biologically-active matrix-derived growth factors. These results support a model in which osteoclastic secretion of protons demineralizes bone, causing release of growth factors from bone matrix. Normally these are largely degraded, with collagen, in the resorptive hemivacuole and during transcytosis to the basal surface of the osteoclast, but in the presence of CathK inhibitor they are released intact, and so might augment bone formation.


Heterogeneity of subsets in glioblastoma mediated by Smad3 palmitoylation.
Glioblastoma (GBM) is the most common and deadly of the primary intracranial tumors and is comprised of subsets that show plasticity and marked heterogeneity, contributing to the lack of success in genomic profiling to guide development of precision medicine for these tumors. In this study, a mutation in isocitrate dehydrogenase 1 was found to suppress the transforming growth factor-beta signaling pathway and E2F4 interacted with Smad3 to inhibit expression of mesenchymal markers. However, palmitoylation of Smad3 mediated by palmitoyltransferase ZDHHC19 promoted activation of the transforming growth factor-beta signaling pathway, and its interaction with EP300 promoted expression of mesenchymal markers in the mesenchymal subtype of GBM. Smad3 and hypoxia-inducible factor 1-alpha may be important molecular targets for treatment of glioma because they appear to coordinate the basic aspects of cancer stem cell biology.


G-protein inhibition profile of the reported Gq/11 inhibitor UBO-QIC.
UBO-QIC (FR900359) is the only currently available Gq/11 protein inhibitor. However, its characterization has not been published, and we thus set out to do this. Gi, Gs and Gq protein-mediated responses were assessed utilizing endogenous or heterologously expressed receptors in Chinese hamster ovary cells. UBO-QIC, at 1 μM, was an effective inhibitor of the Gq-mediated responses, but was inactive at Gi- and Gs-mediated responses. Gq/11 and G16 responses were additionally compared in HEL92.1.7 cells, showing inhibition of Gq/11 responses. However, UBO-QIC also appeared to inhibit G16. Further studies are required to establish its profile with respect to the different Gq-family proteins.


Carbapenem-resistant Pseudomonas aeruginosa: factors influencing multidrug-resistant acquisition in non-critically ill patients.
A cohort study was carried out on hospitalized adult non-critically ill patients (January 2003-December 2004) to identify factors associated with the acquisition of multidrug-resistant Pseudomonas aeruginosa (MDR-PA). A total of 246 non-critically patients were included, 162 (66%) who revealed MDR-PA in the first isolate and 84 (34%) who had carbapenem-resistant P. aeruginosa (CR-PA) isolates. Multivariate analysis identified nosocomial acquisition (odds ratio [OR] 2.7, 95% confidence interval [CI] 1.1-6.3), urinary catheter (OR 2.1, 95%CI 1.1-4.3), and the prior use of fluoroquinolones (OR 2.6, 95%CI 1.0-6.7) as independent risk factors associated with MDR-PA acquisition. Our results show that antibiotics, most notably, fluoroquinolones, may play a major role in the emergence of MDR-PA.