Spanish validation of Charlson index applied to prostate cancer.
Comorbidity assessment is essential in the triage of care for men with prostate cancer (PC). The aim of this study was to validate the Spanish version of the revised Charlson index (RCI) in PC. 731 PC patients diagnosed from 1993 to 2008 were referred to our Radiation Oncology Department. The RCI classified patients into four categories RCI 0, RCI 1-2, RCI 3-4, and RCI 5 and higher. The Kaplan-Meier method and Cox proportional hazards modeling were used. We also analyzed the median age of patients who remained alive at the last control and those who died due to non-prostate cancer comorbidities. 636 patients were included median age: 70 years (44-85). The mean follow-up was 153.62 months, (6-288 months). Distribution of the D'Amico risk classification was 21%, 38.2%, and 40.8% for low, intermediate, and high risk, respectively. The RCI distribution categories were: 303 (46.7%) RCI 0, 102 (16%) RCI 1-2, 131 (20.6%) RCI 3-4, and 100 (15.7%) RCI 5 and higher. The probability of non-cause-specific mortality at 5 and 10 years was 2. 4% and 11.25% RCI 0, 3 and 14.1% RCI 1-2, 5.7% and 22.1% RCI 3-4, and 47% and 92% (RCI 5 and higher). The median age in the last control in patients alive or who had died by non-PC causes was 82.81 years (55.27-102). The RCI may be used to aid medical decision making in older Spanish men with PC, especially in those with a high RCI 5 and higher.


Topical all-trans-retinoic acid prevents corticosteroid-induced skin atrophy without abrogating the anti-inflammatory effect.
We tested the ability of all-trans-retinoic acid to prevent corticosteroid-induced skin atrophy without lessening the anti-inflammatory effect of the steroids. Histologic study and skin-fold thickness in hairless mice treated topically with various steroids, followed by topical all-trans-retinoic acid, were used to measure prevention of atrophy. By both assessments, all-trans-retinoic acid prevented atrophy. Noninterference with the anti-inflammatory property of steroids was tested in a phorbol ester-induced mouse ear edema model and by histologic assessment of croton oil-induced inflammation of mouse dermis. We found that all-trans-retinoic acid did not interfere with steroid suppression of either edema or dermal inflammation. Thus all-trans-retinoic acid was effective in preventing steroid-induced atrophy without affecting the steroid's anti-inflammatory property.


Mutagenicity, sister chromatid exchange inducibility and in vitro cell transforming ability of particulates from Athens air.
Airborne particulates were collected over a period of twelve months by the use of Hi-Vol samplers in the basin of Athens, Greece. N-Hexane extracts were tested in a battery of in vitro tests for their ability to induce mutation in bacteria as well as mutation, sister chromatid exchange and morphological transformation in cultured mammalian cells. Positive results were found for mutagenicity with Salmonella strain TA98 in the Ames assay, for sister chromatid exchange induction in CHO cells and for transformation in BALB/c 3T3 cells in culture. They also showed weak non-dose-related induction of ouabain resistance in BALB/c 3T3 cells. The contribution of oxidizing and nitrating agents found in the Athens atmosphere, together with sunlight UV irradiation in the formation of direct acting mutagens and potential carcinogens from ambient polycyclic aromatic hydrocarbons, is suggested.


Double edge redox-implications for the interaction between endogenous thiols and copper ions: In vitro studies.
The present study investigated the redox-consequences of the interaction between various endogenous thiols (RSH)-glutathione, cysteine, homocysteine, gamma-glutamyl-cysteine, and cysteinyl-glycine- and Cu(2+) ions, in terms of their free radical-scavenging, ascorbate-oxidizing and O2(*-)-generating properties of the resulting mixtures. Upon a brief incubation (3-30 min) with Cu(2+), the free radical-scavenging properties (towards ABTS(*)(+) and DPPH(*)) and thiol-titratable groups of the RSH added to the mixtures decreased significantly. Remarkably, both effects were only partial, even in the presence of a large molar Cu(2+)-excess, and were unaffected despite increasing the incubation time. At equimolar concentrations, the RSH/Cu(2+) mixtures led to the formation of (EPR paramagnetic) Cu(II)-complexes that were time-stable and ascorbate-reducible, but redox-inactive towards oxygen. In turn, at a slight molar thiol-excess (3:1), the mixtures resulted in the formation of time-stable Cu(I)-complexes (EPR silent) that were unreactive towards ascorbate and oxygen. The only exception was seen for the thiol, glutathione, whose mixture with Cu(2+) mixture displayed a O2(*-)-generating capacity (cytochrome c- and lucigenin-reduction). The data indicate that, depending on their molar ratio, the interaction between Cu(2+) and the tested thiols would give place to mixtures containing either: (i) time-stable and ascorbate-reducible Cu(II)-complexes which display free radical-scavenging properties, or (ii) time-stable but redox-inactive towards oxygen Cu(I)-complexes. Among the latter, the only exception was that of glutathione.


Psychomotor retardation is linked to frontal alpha asymmetry in major depression.
Psychomotor disturbances are a main clinical feature of major depressive disorder (MDD) but little is known about their EEG signature. One of the most replicated EEG findings in MDD is resting frontal asymmetry in the alpha band (FAA), which is thought to be a correlate of withdrawal behavior and reduced approach motivation. The purpose of this study was to assess psychomotor alterations, alpha band power, FAA and investigate the association between them. 20 MDD patients and 19 healthy subjects were enrolled. Alpha power and FAA scores were calculated from a resting state EEG. Wrist actigraphy was recorded from the non-dominant arm for 24 h and activity level scores (AL) were extrapolated from the wakeful periods. MDD patients had a left-lateralized frontal alpha activity and lower AL scores when compared to healthy subjects. A significant correlation was found between mean FAA and AL scores. A negative covariance between power in the lower alpha range and AL scores over the motor cortex bilaterally was detected. Relatively small sample size. Patients were pharmacologically treated with antidepressants. This study replicates the finding of left-lateralized FAA and lower AL scores in MDD patients, and establishes the first evidence of significant correlations between alpha power, FAA scores and measures of motor activity, which may be interpreted as an expression of impaired motivational drive in MDD.


Actions of alinidine and AQ-A 39 on rate and contractility of guinea pig atria during beta-adrenoceptor stimulation.
We studied two new agents, alinidine (St 567, 2-[N-allyl-N-(2,6-dichlorophenyl)-amino]-2-imidazoline) and AQ-A 39 (5,6-dimethoxy-2-[3[[alpha-(3,4-dimethoxy)-phenylethyl] methylamino]propyl]phthalimidine), in isolated guinea pig atria with respect to their specificity to decrease heart rate but not contractility during beta-adrenoceptor stimulation. Spontaneous electrical activity in sinoatrial node preparations was increased by perfusion with isoprenaline (0.1 micrograms/ml); addition of alinidine (3 micrograms/ml), AQ-A 39 (3 micrograms/ml), or propranolol (0.03-0.3 micrograms/ml) reduced sinus rate to the control values. The same concentrations of these drugs were tested in electrically driven (1 Hz) left atria. The positive inotropic effect of isoprenaline was not affected by alinidine and AQ-A 39; however, it was markedly reduced or abolished by propranolol. The experiments, therefore, demonstrated the bradycardic specificity of alinidine and AQ-A 39 under conditions of beta-adrenoceptor stimulation. A cumulative concentration-response curve for the positive chronotropic effect of isoprenaline was established in spontaneously beating atria. AQ-A 39, 1 and 10 micrograms/ml, reduced the control sinus rate but did not affect the following concentration-response curve of isoprenaline. A similar result was published earlier for alinidine and excludes an interaction of both drugs with isoprenaline on a possible subgroup of selective beta-adrenoceptors in the sinoatrial node.


Up-regulation of inducible nitric oxide synthase and nitric oxide in Helicobacter pylori-infected human gastric epithelial cells: possible role of interferon-gamma in polarized nitric oxide secretion.
Nitric oxide (NO) generated by nitric oxide synthase (NOS) is known to be an important modulator of the mucosal inflammatory response. In this study, we questioned whether Helicobacter pylori infection could up-regulate the epithelial cell inducible NOS (iNOS) gene expression and whether NO production could show polarity that can be regulated by immune mediators. Human gastric epithelial cell lines were infected with H. pylori, and the iNOS mRNA expression was assessed by quantitative RT-PCR. NO production was assayed by determining nitrite/nitrate levels in culture supernatants. To determine the polarity of NO secretion by the H. pylori-infected epithelial cells, Caco-2 cells were cultured as polarized monolayers in transwell chambers, and NO production was measured. iNOS mRNA levels were significantly up-regulated in the cells infected with H. pylori, and expression of iNOS protein was confirmed by Western blot analysis. Increased NO production in the gastric epithelial cells was seen as early as 18 hours postinfection, and reached maximal levels by 24 hours postinfection. The specific MAP kinase inhibitors decreased H. pylori-induced iNOS and NO up-regulation. After H. pylori infection of polarized epithelial cells, NO was released predominantly into the apical compartment, and IL-8 was released predominantly into basolateral compartment. The addition of IFN-gamma to H. pylori-infected polarized epithelial cells showed a synergistically higher apical and basolateral NO release. These results suggest that apical NO production mediated by MAP kinase in H. pylori-infected gastric epithelial cells may influence the bacteria and basolateral production of NO and IL-8 may play a role in the tissue inflammation.


Exploring the motivations behind misreporting self-measured blood glucose in adolescents with type 1 diabetes - a qualitative study.
Despite advances in diabetes management, the reporting and self-monitoring of blood glucose (SMBG) remains fundamental. While previous work has established that the misreporting of SMBG to family and medical professionals is surprisingly common, the motivations behind this behaviour have never been examined. We aimed to investigate the motivations behind misreporting of SMBG in adolescents with type 1 diabetes (T1DM). Fifteen semi-structured interviews were conducted with adolescents (aged 12-19 inclusive) with T1DM recruited through diabetes clinics across the Otago/Southland region of New Zealand from November 2015 to January 2016. These were transcribed and content analysis performed to identify themes and subthemes in misreporting behaviour. The mean age of participants was 15.7 years, 60 % were male, with 67 % using multiple daily insulin injections, and 33 % on insulin pumps. Their median HbA1c was 84 mmol/mol, range 52-130. Misreporting behaviour was described for both electronic pump records and written logbooks, as well as verbally. Multiple motivations for misreporting were given, spanning three major themes: Achieving potential benefits; the avoidance of negative consequences; and the avoidance of worry/concern (in self or in others). The main suggestion of participants to reduce misreporting behaviour was to reduce the negative reactions of others to suboptimal blood glucose readings. Electronic, written, and verbal SMBG misreporting remains common. This study provides deeper insight into the motivations leading to this behaviour in adolescents, suggesting that further understanding and attention to this aspect of adherence may lead to improvements not only in glycaemic control and safety, but also to the psychological wellbeing of those with T1DM.


Associations between immediate postpartum long-acting reversible contraception and short interpregnancy intervals.
We aimed to evaluate the rates of short interpregnancy interval pregnancies and deliveries among women who receive immediate postpartum LARC. We conducted a retrospective cohort study of all women who delivered at Montefiore Medical Center between January 2015 and June 2016 (N = 9561). In this cohort, we identified all repeat deliveries and pregnancies within 18 months of the initial delivery. Using logistic regression models, we compared rates of short interpregnancy interval delivery and pregnancy among recipients of an immediate postpartum IUD, immediate postpartum implant, and no immediate postpartum LARC, adjusting for covariates including patient age, mode of delivery, socioeconomic status, and race. In our cohort, 12.9% of patients received immediate postpartum LARC. The rates of short interpregnancy interval delivery were 3.3% (N = 259/7833) among patients who did not receive immediate postpartum LARC, 1% (N = 6/595) among immediate postpartum IUD recipients, and 0.4% (N = 2/562) among immediate postpartum implant recipients. The rates of short interpregnancy interval pregnancy were 13.8% (N = 1082/7833) among patients who did not receive immediate postpartum LARC, 7.4% (N = 44/595) among immediate postpartum IUD recipients, and 5.2% (N = 29/562) among immediate postpartum implant recipients. Both recipients of immediate postpartum IUDs and immediate postpartum implants had lower rates of short interpregnancy interval delivery and pregnancy compared to patients who did not receive immediate postpartum LARC. This study confirms that women who received immediate postpartum IUDs and implants have lower rates of short interpregnancy interval pregnancies. Making immediate postpartum LARC widely available is a promising public health approach to help women achieve a longer interpregnancy interval.


Author Correction: When the statistical MMN meets the physical MMN.
An amendment to this paper has been published and can be accessed via a link at the top of the paper.


Doppler atrial shunt flow patterns in patients with secundum atrial septal defect: determinants, limitations, and pitfalls.
Fifteen patients with uncomplicated secundum atrial septal defect underwent studies with real-time color-coded two dimensional flow imaging, pulsed Doppler echocardiographic examination, and simultaneous pressure recordings from the left and right atrium to determine the flow-pressure dynamics of the atrial shunt flow. In all 15 patients both the color flow mapping and pulsed Doppler studies revealed that the shunt flow was mainly from left to right, occurring both during ventricular systole and diastole. It started in early systole, reached a peak in late systole to early diastole, and lasted throughout diastole with an accentuation in late diastole during atrial contraction. The amplitude of the flow velocity, the direction, and the magnitude of the shunt flow, however, changed from phase to phase during the cardiac cycle. It correlated well with the phasic variation of the interatrial pressure difference, which usually revealed a peak pressure gradient that occurred in early systole between the x descent and v wave and during the period of v wave and a wave of the left atrial pressure tracing. Right to left shunt was not detected in any of the 15 patients by color flow mapping studies. A minor reversal of the shunt flow, however, was frequently detected at the beginning of ventricular systole and sometimes also in the middle of diastole by pulsed Doppler echocardiography. The reversal of shunt flow correlated with the minor reversal of pressure gradient that occurred during the z point, x descent, and y descent of the left atrial pressure tracing. In conclusion, left to right shunt flow occurs both during ventricular systole and diastole in uncomplicated secundum atrial septal defect.(ABSTRACT TRUNCATED AT 250 WORDS)


Limitations of cardiothoracic ratio derived from chest radiographs to predict real heart size: comparison with magnetic resonance imaging.
Cardiothoracic ratio (CTR) in chest radiographs is still widely used to estimate cardiac size despite the advent of newer imaging techniques. We hypothesise that a universal CTR cut-off value of 50% is a poor indicator of cardiac enlargement. Our aim was to compare CTR with volumetric and functional parameters derived from cardiac magnetic resonance imaging (MRI). 309 patients with a chest radiograph and cardiac MRI acquired within a month were reviewed to assess how CTR correlates with multiple cardiac MRI variables: bi-ventricular EDV (absolute and indexed to body surface area), EF, indexed total heart volume and bi-atrial areas. In addition, we have also determined CTR accuracy by creating multiple ROC curves with the described variables. All cardiac MRI variables correlate weakly but statistically significantly with CTR. This weak correlation is explained by a substantial overlap of cardiac MRI parameters in patients with normal and increased CTR. For all variables, CTR was only mildly to moderately better than a chance to discriminate cardiac enlargement (AUC 0.6-0.7). Large CTR values (> 55%) are specific but not sensitive, while low CTR values (< 45%) are sensitive but not specific. Values in between are not sensitive nor specific. CTR correlates weakly with true chamber size assessed by gold standard cardiac MRI and has a weak discriminatory power. Thus, clinical decisions based on intermediate CTRs (45-55%) should be avoided. Large CTRs (> 55%) are likely indicative of true heart chamber enlargement. Low CTRs (< 45%) are likely indicative of normal heart size.


Neuroprotection by stem cell factor in rat cortical neurons involves AKT and NFkappaB.
Stem cell factor (SCF) is a highly expressed cytokine in the central nervous system. In the present study, we demonstrate a neuroprotective role for SCF and its tyrosine kinase receptor, c-kit, against camptothecin-induced apoptosis and glutamate excitotoxicity in rat cortical neurons. This protection was blocked by pharmacological or molecular inhibition of either the MEK/ERK or PI3K/Akt signaling pathways. The importance of these pathways was further confirmed by the activation of both ERK, in a MEK-dependent manner, and Akt, via PI3K. Activation of Akt increased the binding of the p50 and p65 subunits of NFkappaB, which was also important for neuroprotection. Akt inhibition prevented NFkappaB binding, suggesting a role for Akt in SCF-induced NFkappaB. Pharmacological inhibition of NFkappaB or dominant negative IkappaB also prevented neuroprotection by SCF. SCF up-regulated the anti-apoptotic genes, bcl-2 and bcl-xL in an NFkappaB-dependent manner. Together, these findings demonstrate a neuroprotective role for SCF in cortical neurons, an effect that was mediated by Akt and ERK, as well as NFkappaB-mediated gene transcription. SCF represents a novel therapeutic target in the treatment of neurodegenerative disease.


Impact of COVID-19 lockdown on glycemic control in type 1 diabetes.
The COVID-19 pandemic has forced governments to impose lockdown policies, thus impacting patients with chronic diseases, such as type 1 diabetes. The aim of this study was to evaluate the impact of lockdown on glycemic control in type 1 diabetes patients. We retrospectively evaluated patients using a continuous subcutaneous insulin infusion device during the nationwide lockdown. Children and adolescents aged 2-18 years followed up at the Pediatric Endocrinology Unit of Hospitalar São João in Portugal were included in the study. We collected data on the age, weight, insulin doses, and glycemic control of the patients before and after the restrictions. The study included 100 patients, 59 males, with a mean age of 12.5 years. Baseline data showed a suboptimal glycemic control with a median HbA1c of 7.9%. The lockdown was associated with an increase in the body mass index (BMI) of all patients (p = 0.009), particularly girls and older teenagers. Metabolic control deteriorated in the 10-13 age group (p = 0.03), with a 0.4% increase in HbA1c. To date, this is the largest study on the impact of lockdown on type 1 diabetes in patients using an insulin pump. The results highlight the importance of physical activity, parental supervision, and continuation of healthcare assistance through telemedicine in young individuals with type 1 diabetes.


Enhanced resistance to Bacillus thuringiensis Cry1Ac toxin mediated by the activation of prophenoloxidase in a cosmopolitan pest.
Plutella xylostella has evolved resistance to Bacillus thuringiensis Cry1Ac toxin over a long evolutionary period. Enhanced immune response is an important factor in insect resistance to a variety of insecticides, and whether phenoloxidase (PO), an immune protein, is involved in resistance to Cry1Ac toxin in P. xylostella remains unclear. Here, spatial and temporal expression patterns showed that prophenoloxidase (PxPPO1 and PxPPO2) in the Cry1S1000-resistant strain was more highly expressed in eggs, 4th instar, head, and hemolymph than those in G88-susceptible strain. The results of PO activity analysis showed that after treatment with Cry1Ac toxin PO activity was about 3 times higher than that before treatment. Furthermore, knockout of PxPPO1 and PxPPO2 significantly increased the susceptibility to Cry1Ac toxin. These findings were further supported by the knockdown of Clip-SPH2, a negative regulator of PO, which resulted in increased PxPPO1 and PxPPO2 expression and Cry1Ac susceptibility in the Cry1S1000-resistant strain. Finally, the synergistic effect of quercetin showed that larval survival decreased from 100 % to <20 % compared to the control group. This study will provide a theoretical basis for the analysis of immune-related genes (PO) genes involved in the resistance mechanism and pest control of P. xylostella.


A CCL24-dependent pathway augments eosinophilic airway inflammation in house dust mite-challenged Cd163(-/-) mice.
CD163 is a macrophage scavenger receptor with anti-inflammatory and pro-inflammatory functions. Here, we report that alveolar macrophages (AMΦs) from asthmatic subjects had reduced cell-surface expression of CD163, which suggested that CD163 might modulate the pathogenesis of asthma. Consistent with this, house dust mite (HDM)-challenged Cd163(-/-) mice displayed increases in airway eosinophils and mucous cell metaplasia (MCM). The increased airway eosinophils and MCM in HDM-challenged Cd163(-/-) mice were mediated by augmented CCL24 production and could be reversed by administration of a neutralizing anti-CCL24 antibody. A proteomic analysis identified the calcium-dependent binding of CD163 to Dermatophagoides pteronyssinus peptidase 1 (Der p1). Der p1-challenged Cd163(-/-) mice had the same phenotype as HDM-challenged Cd163(-/-) mice with increases in airway eosinophils, MCM and CCL24 production, while Der p1 induced CCL24 secretion by bone marrow-derived macrophages (BMMΦs) from Cd163(-/-) mice, but not BMMΦs from wild-type (WT) mice. Finally, airway eosinophils and bronchoalveolar lavage fluid CCL24 levels were increased in Der p1-challenged WT mice that received adoptively transferred AMΦ's from Cd163(-/-) mice. Thus, we have identified CD163 as a macrophage receptor that binds Der p1. Furthermore, we have shown that HDM-challenged Cd163(-/-) mice have increased eosinophilic airway inflammation and MCM that are mediated by a CCL24-dependent mechanism.


Pulsatile cervical swelling (dilated carotid artery) associated with persistent primitive trigeminal artery. Case report.
A 39-year-old woman was admitted to the department of surgery due to a left-sided pulsatile cervical swelling. Angiography showed that the pulsatile swelling was due to enlargement of the internal carotid artery, and revealed a persistent left-sided primitive trigeminal artery. The clinical significance of this persistent carotid-basilar anastomosis is reviewed, and its possible pathogenetic relationship with the pulsatile cervical swelling is discussed.


The postprandial secretion of peptide YY1-36 and 3-36 in obesity is differentially increased after gastric bypass versus sleeve gastrectomy.
Peptide tyrosine tyrosine (PYY) exists as two species, PYY1-36 and PYY3-36 , with distinct effects on insulin secretion and appetite regulation. The detailed effects of bariatric surgery on PYY1-36 and PYY3-36 secretion are not known as previous studies have used nonspecific immunoassays to measure total PYY. Our objective was to characterize the effect of sleeve gastrectomy (SG) and Roux-en-Y gastric bypass (RYGB) on fasting and postprandial PYY1-36 and PYY3-36 secretion using a newly developed liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay. Observational study in 10 healthy nonobese volunteers and 30 participants with obesity who underwent RYGB (n = 24) or SG (n = 6) at the Imperial Weight Centre [NCT01945840]. Participants were studied using a standardized mixed meal test (MMT) before and 1 year after surgery. The outcome measures were PYY1-36 and PYY3-36 concentrations. Presurgery, the fasting and postprandial levels of PYY1-36 and PYY3-36 were low, with minimal responses to the MMT, and these did not differ from healthy nonobese volunteers. The postprandial secretion of both PYY1-36 and PYY3-36 at 1 year was amplified after RYGB, but not SG, with the response being significantly higher in RYGB compared with SG. There appears to be no difference in PYY secretion between nonobese and obese volunteers at baseline. At 1 year after surgery, RYGB, but not SG, is associated with increased postprandial secretion of PYY1-36 and PYY3-36 , which may account for long-term differences in efficacy and adverse effects between the two types of surgery.


Progression elevated gene-3 promoter (PEG-Prom) confers cancer cell selectivity to human polynucleotide phosphorylase (hPNPase(old-35))-mediated growth suppression.
The poor prognosis of pancreatic cancer patients using currently available therapies mandates novel therapeutics that combine anti-neoplastic potency with toxicity-minimizing cancer specificity. Employing an overlapping pathway screen to identify genes exhibiting coordinated expression as a consequence of terminal cell differentiation and replicative senescence, we identified human polynucleotide phosphorylase (hPNPase(old-35)), a 3',5'-exoribonuclease that exhibits robust growth-suppressing effects in a wide spectrum of human cancers. A limitation to the anti-neoplastic efficacy of hPNPase(old-35) relates to its lack of cancer specificity. The promoter of Progression Elevated Gene-3 (PEG-Prom), discovered in our laboratory via subtraction hybridization in a transformation progression rodent tumor model functions selectively in a diverse array of human cancer cells, with limited activity in normal cells. An adenovirus constructed with the PEG-Prom driving expression of hPNPase(old-35) containing a C-terminal Hemaglutinin (HA)-tag (Ad.PEG.hPNPase(old-35)) was shown to induce robust transgene expression, growth suppression, apoptosis, and cell-cycle arrest in a broad panel of pancreatic cancer cells, with minimal effects in normal immortalized pancreatic cells. hPNPase(old-35) expression correlated with arrest in the G(2)/M phase of the cell cycle and up-regulation of the cyclin-dependent kinase inhibitors (CDKI) p21(CIP1/WAF-1/MDA-6) and p27(KIP1). In a nude mouse xenograft model, Ad.PEG.hPNPase(old-35) injections effectively inhibited growth of human pancreatic cancer cells in vivo. These findings support the potential efficacy of combining a cancer-specific promoter, such as the PEG-Prom, with a novel anti-neoplastic agent, such as hPNPase(old-35), to create a potent, targeted cancer therapeutic, especially for a devastating disease like pancreatic cancer.


Simulations of octapeptin-outer membrane interactions reveal conformational flexibility is linked to antimicrobial potency.
The octapeptins are lipopeptide antibiotics that are structurally similar to polymyxins yet retain activity against polymyxin-resistant Gram-negative pathogens, suggesting they might be used to treat recalcitrant infections. However, the basis of their unique activity is unclear because of the difficulty in generating high-resolution experimental data of the interaction of antimicrobial peptides with lipid membranes. To elucidate these structure-activity relationships, we employed all-atom molecular dynamics simulations with umbrella sampling to investigate the conformational and energetic landscape of octapeptins interacting with bacterial outer membrane (OM). Specifically, we examined the interaction of octapeptin C4 and FADDI-115, lacking a single hydroxyl group compared with octapeptin C4, with the lipid A-phosphoethanolamine modified OM of Acinetobacter baumannii Octapeptin C4 and FADDI-115 both penetrated into the OM hydrophobic center but experienced different conformational transitions from an unfolded to a folded state that was highly dependent on the structural flexibility of their respective N-terminal fatty acyl groups. The additional hydroxyl group present in the fatty acyl group of octapeptin C4 resulted in the molecule becoming trapped in a semifolded state, leading to a higher free energy barrier for OM penetration. The free energy barrier for the translocation through the OM hydrophobic layer was ∼72 kcal/mol for octapeptin C4 and 62 kcal/mol for FADDI-115. Our results help to explain the lower antimicrobial activity previously observed for octapeptin C4 compared with FADDI-115 and more broadly improve our understanding of the structure-function relationships of octapeptins. These findings may facilitate the discovery of next-generation octapeptins against polymyxin-resistant Gram-negative 'superbugs.'


Kinetics of indium-III-labeled eosinophils in two patients with eosinophilia.
The kinetics of IIIIn-labeled eosinophils was evaluated in two cases of eosinophilia by means of external and blood cell counting. In both cases, the labeled blood eosinophils decreased rapidly for a few h after injection and then increased progressively up to the 20th and the 12th h, respectively. The changes of radioactivity with time in the liver, spleen and lungs correlated with that in the blood eosinophils. However, in both cases radioactivity of the thoracic spine increased rapidly for a few h after injection of the labeled eosinophils and subsequently decreased. This result suggests that some of the eosinophils recirculate between the marginal pool of marrow and the intra vascular space.


A new system to detect native microaggregates of platelets in vivo, with a novel platelet aggregometer employing laser light scattering.
Circulating platelets are susceptible to various stimuli in vessels, and a certain portion of platelets may thereby form native microaggregates (NMAs), which can be visualized by a bother scanning electron microscopy (SEM). We developed an easy method to evaluate the existence and extent of NMAs and the subsequent spontaneous platelet aggregation (SPA) induced by not any agonist but stirring force with a novel platelet aggregometer (PA-200; Kowa, Tsukuba, Japan) employing a particle-counting method based on laser light scattering. In this system, platelet-rich plasma obtained by a delicate centrifugation was infused in a cuvette to replace the content and to avoid stirring shear. An insertion attachment was designed in a coaxial shape to maintain a constant sample volume. Our data indicated that NMAs, tentatively defined as a "small" division by their light scattering (25-400 of the intensity threshold), could be detected by not only a complicated SEM but also by our simple system with the PA-200 in a few minutes. No correlation was found in the comparison of the existence of NMAs and the SPA generation, in aliquot platelets, respectively. These results suggested that NMAs and SPA might be mutually different cluster and have meanings as representations of platelet status.


Use of fluorescence in situ hybridization to predict response to bacillus Calmette-Guérin therapy for bladder cancer: results of a prospective trial.
No reliable methods currently exist to predict patient response to intravesical immunotherapy with bacillus Calmette-Guérin given after transurethral resection for high risk nonmuscle invasive bladder cancer. We initiated a prospective clinical trial to determine whether fluorescence in situ hybridization results during bacillus Calmette-Guérin immunotherapy can predict therapy failure. Candidates for standard of care bacillus Calmette-Guérin were offered participation in a clinical trial. Fluorescence in situ hybridization was performed before bacillus Calmette-Guérin, and at 6 weeks, 3 months and 6 months during bacillus Calmette-Guérin therapy with maintenance. Cox proportional hazards regression was used to assess the relationship between fluorescence in situ hybridization results and tumor recurrence or progression. The Kaplan-Meier product limit method was used to estimate recurrence-free and progression-free survival. A total of 126 patients participated in the study. At a median followup of 24 months 31% of patients had recurrent tumors and 14% experienced disease progression. Patients who had positive fluorescence in situ hybridization results during bacillus Calmette-Guérin therapy were 3 to 5 times more likely than those who had negative fluorescence in situ hybridization results to experience recurrent tumors and 5 to 13 times more likely to have disease progression (p <0.01). The timing of positive fluorescence in situ hybridization results also affected outcomes. For example, patients with a negative fluorescence in situ hybridization result at baseline, 6 weeks and 3 months demonstrated an 8.3% recurrence rate compared to 48.1% for those with a positive result at all 3 points. Fluorescence in situ hybridization results can identify patients at risk for tumor recurrence and progression during bacillus Calmette-Guérin immunotherapy. This information may be used to counsel patients about alternative treatment strategies.


Gene expression profiling in Salmonella Choleraesuis-infected porcine lung using a long oligonucleotide microarray.
Understanding the transcriptional response to pathogenic bacterial infection within food animals is of fundamental and applied interest. To determine the transcriptional response to Salmonella enterica serovar Choleraesuis (SC) infection, a 13,297-oligonucleotide swine array was used to analyze RNA from control, 24-h postinoculation (hpi), and 48-hpi porcine lung tissue from pigs infected with SC. In total, 57 genes showed differential expression (p < 0.001; false discovery rate = 12%). Quantitative real-time PCR (qRT-PCR) of 61 genes was used to confirm the microarray results and to identify pathways responding to infection. Of the 33 genes identified by microarray analysis as differentially expressed, 23 were confirmed by qRT-PCR results. A novel finding was that two transglutaminase family genes (TGM1 and TGM3) showed dramatic increases in expression postinoculation; combined with several other apoptotic genes, they indicated the induction of apoptotic pathways during SC infection. A predominant T helper 1-type immune response occurred during infection, with interferon gamma (IFNG) significantly increased at 48 hpi. Genes induced by IFNs (GBP1, GBP2, C1S, C1R, MHC2TA, PSMB8, TAP1, TAP2) showed increased expression during porcine lung infection. These data represent the first thorough investigation of gene regulation pathways that control an important porcine respiratory and foodborne bacterial infection.


Low-dose cytosine arabinoside (LD-AraC) vs LD-AraC plus granulocyte/macrophage colony stimulating factor vs LD-AraC plus Interleukin-3 for myelodysplastic syndrome patients with a high risk of developing acute leukemia: final results of a randomized phase III study (06903) of the EORTC Leukemia Cooperative Group.
In this randomized phase III study of the EORTC Leukemia Cooperative Group, patients with myelodysplastic syndromes (MDS) with 10-30% bone marrow blasts and hematopoietic failure were treated with low-dose cytosine arabinoside (LD-AraC) (2 x 10 mg/m2/day subcutaneously (s.c.) days 1-14) either alone or in combination with rhGM-CSF or interleukin-3 (IL-3) both given s.c. at a dose of 150 microg/day from day 8 to 21. A total of 180 evaluable patients with a median age of 65 years and refractory anemia with an excess of blasts (RAEB, n = 107) or RAEB in transformation (RAEBt, n = 73) were randomized. There were no differences among the three treatment regimens with respect to numbers of courses applied or treatment delays. Hemorrhage occurred in approximately 40% in all arms, whereas infection rates were higher in the granulocyte/macrophage colony stimulating factor (GM-CSF)- or IL3-containing arm. The overall response rate was 38.6% with no statistically significant difference among the three arms. In summary, a substantial proportion of patients had achieved relatively durable responses in all the three arms. No influence of either growth factor was detected on the grade of cytopenia. Thus, the combination of LD-AraC with GM-CSF or IL-3 cannot be recommended for routine use in a high-risk MDS population.


Novel approaches to treatment of hepatocellular carcinoma and hepatic metastases using thermal ablation and thermosensitive liposomes.
Because of the limitations of surgical resection, thermal ablation is commonly used for the treatment of hepatocellular carcinoma and liver metastases. Current methods of ablation can result in marginal recurrences of larger lesions and in tumors located near large vessels. This review presents a novel approach for extending treatment out to the margins where temperatures do not provide complete treatment with ablation alone, by combining thermal ablation with drug-loaded thermosensitive liposomes. A history of the development of thermosensitive liposomes is presented. Clinical trials have shown that the combination of radiofrequency ablation and doxorubicin-loaded thermosensitive liposomes is a promising treatment.


[Antiischemic efficacy of trimetazidine in patients with intermittent claudication and effort angina].
Male patients (n=42) with atherosclerosis obliterans and stage IIA-III ischemia of lower extremities combined with class II-III angina pectoris were divided into 2 groups. Patients of group 1 (n=21) were given simvastatin (10-20 mg/day), clopidogrel (75 mg/day) and trimetazidine (60 mg/day) for 6 months, those of group 2 - simvastatin and clopidogrel also for 6 months. Statistically significant decreases of frequency of attacks of angina, nitroglycerine consumption and increases of walking distance occurred only in group 1.


Comparative proteome and serum analysis identified FSCN1 as a marker of abiraterone resistance in castration-resistant prostate cancer.
Abiraterone (Abi) is an androgen receptor signaling inhibitor that significantly improves patients' life expectancy in metastatic prostate cancer (PCa). Despite its beneficial effects, many patients have baseline or acquired resistance against Abi. The aim of this study was to identify predictive serum biomarkers for Abi treatment. We performed a comparative proteome analysis on three Abi sensitive (LNCaPabl, LAPC4, DuCaP) and resistant (LNCaPabl-Abi, LAPC4-Abi, DuCaP-Abi) PCa cell lines using liquid chromatography tandem mass spectrometry (LC-MS/MS) technique. Two bioinformatic selection workflows were applied to select the most promising candidate serum markers. Serum levels of selected proteins were assessed in samples of 100 Abi-treated patients with metastatic castration-resistant disease (mCRPC) using ELISA. Moreover, FSCN1 serum concentrations were measured in samples of 69 Docetaxel (Doc) treated mCRPC patients. Our proteome analysis identified 68 significantly, at least two-fold upregulated proteins in Abi resistant cells. Using two filtering workflows four proteins (AMACR, KLK2, FSCN1 and CTAG1A) were selected for ELISA analyses. We found high baseline FSCN1 serum levels to be significantly associated with poor survival in Abi-treated mCRPC patients. Moreover, the multivariable analysis revealed that higher ECOG status (>1) and high baseline FSCN1 serum levels (>10.22 ng/ml by ROC cut-off) were independently associated with worse survival in Abi-treated patients (p < 0.001 and p = 0.021, respectively). In contrast, no association was found between serum FSCN1 concentrations and overall survival in Doc-treated patients. Our analysis identified baseline FSCN1 serum levels to be independently associated with poor survival of Abi-treated, but not Doc-treated mCRPC patients, suggesting a therapy specific prognostic value for FSCN1.


Melatonin alleviates cisplatin-induced mice spermatogenesis defects.
Cisplatin (CDDP) is a chemotherapeutic drug that is used to treat many different types of tumors. However, it also has significant adverse effects on male reproduction, which are partially mediated oxidative damage. Melatonin (MLT) is a promising antioxidant that can be used for reproductive protection. In this paper, we investigated the effect of CDDP on spermatogenesis, as well as MLT's potential role in reproductive protection. CDDP (5 mg/kg BW) significantly reduced male mice testosterone levels and decreased sperm vitality and progressive motility. Additionally, a lower percentage of stage VII and VIII seminiferous tubules were observed in CDDP-treated mice. MLT administration significantly alleviated CDDP-induced testicular damages, CDDP-induced lowered male fertility in vivo, and enhanced in vitro embryonic development of two cells and blastocysts. These changes may be due to CDDP-mediated spermatogenesis defects in germ cell and Leydig cell proliferation, which are reflected in abnormal PCNA, SYCP3, and CYP11A1 expression levels and can be improved by MLT. CDDP treatment significantly decreased the total antioxidant capacity (TAC), as well as SOD and GSH levels, and increased MDA levels in mice testis, leading to increased apoptosis of germ cells and increased BAX/BCL2 ratios in mice testis. MLT treatment may reduce germ cell apoptosis by reducing oxidative damage in mice testis. This study demonstrated that CDDP affects sperm fertility by altering germ cell and Leydig cell proliferation via increased oxidative damage and that MLT can attenuate these damages. Our work provides potential information for further research on the toxic effects of CDDP and the protective effects of MLT on male reproduction.


Molecular Dynamics Simulations of Kir2.2 Interactions with an Ensemble of Cholesterol Molecules.
Cholesterol is a major regulator of multiple types of ion channels, but the specific mechanisms and the dynamics of its interactions with the channels are not well understood. Kir2 channels were shown to be sensitive to cholesterol through direct interactions with "cholesterol-sensitive" regions on the channel protein. In this work, we used Martini coarse-grained simulations to analyze the long (μs) timescale dynamics of cholesterol with Kir2.2 channels embedded into a model membrane containing POPC phospholipid with 30 mol% cholesterol. This approach allows us to simulate the dynamic, unbiased migration of cholesterol molecules from the lipid membrane environment to the protein surface of Kir2.2 and explore the favorability of cholesterol interactions at both surface sites and recessed pockets of the channel. We found that the cholesterol environment surrounding Kir channels forms a complex milieu of different short- and long-term interactions, with multiple cholesterol molecules concurrently interacting with the channel. Furthermore, utilizing principles from network theory, we identified four discrete cholesterol-binding sites within the previously identified cholesterol-sensitive region that exist depending on the conformational state of the channel-open or closed. We also discovered that a twofold decrease in the cholesterol level of the membrane, which we found earlier to increase Kir2 activity, results in a site-specific decrease of cholesterol occupancy at these sites in both the open and closed states: cholesterol molecules at the deepest of these discrete sites shows no change in occupancy at different cholesterol levels, whereas the remaining sites showed a marked decrease in occupancy.


Biliary Atresia: histopathologic observations and reflections upon its natural history.
Operative liver biopsies and specimens of the extrahepatic ducts and porta hepatis have been studied in 12 cases of biliary atresia. In all cases, the liver showed giant cell transformation and inflammation with mononuclear cells and neutrophils. Most had other features of neonatal hepatitis, including necrosis of hepatocytes. In the intrahepatic bile ducts of all cases but one, the hepatic ducts and glands at the porta hepatis, and in the extrahepatic ducts where epithelium remained, there was degeneration of the epithelium and intramural inflammation. In the ducts at the porta hepatis and in 6 of 8 extrahepatic ducts where epithelium remained, there was extensive mural fibrosis compromising the diameter of the duct lumens. Three cases showed the inflammatory changes distal to sites of closure of the extrahepatic ducts. These findings demonstrate that in biliary atresia, hepatitis, intrahepatic cholangitis, and sclerosing cholangitis of the extrahepatic ducts all interact to produce acquired obstruction to bile flow.


Factors Affecting Outcomes of Craniofacial Clinical Trials.
This study aims to provide a cross-sectional view of craniofacial trials and to identify factors associated with completion, publication, and trial longevity. This is a cross-sectional study of cleft and craniofacial clinical trials registered with ClinicalTrials.gov between September 1999 and April 2020. Predictor variables included funding source, trial design, trial location, number of recruitment sites, and investigator specialty.Study outcomes were completion status, publication status, and trial duration. Univariate comparisons and multivariate regression models were calculated for each outcome. The final sample included 179 clinical trials pertaining to craniofacial care. Nearly all trials were single-center (86.5%), and roughly half of trials were interventional (57.0%) or conducted in the United States (40.5%). No single specialty predominated, although plastic surgery (13.4%) was the most common investigator specialty. The completion rate was 82.7%, the publication rate was 40.8%, and the mean trial duration was 39.1 months. Interventional design (odds ratio = 0.30, P = 0.02) and United States location (odds ratio = 0.15, P < 0.01) were each independently associated with lower odds of trial completion. Trial longevity was independently associated with the National Institute of Health-funding (P < 0.01) and multicenter design (P < 0.01). Craniofacial trials are multidisciplinary and have a high rate of completion. Although most existing trials were conducted at only a single-center, multicenter efforts significantly increased trial longevity without compromising completion and publication rates. Given the diverse array of conditions and lines of inquiry that compose craniofacial care, it is reassuring that collaboration did not negatively affect trial outcomes.


Effect of rapid maxillary expansion on the dimension of the nasal cavity and on nasal air resistance.
Atresia of the maxilla is a transverse skeletal dysplasia, possibly associated with respiratory problems. For its correction, rapid maxillary expansion is a feasible orthodontic process. To evaluate the effect of rapid maxillary expansion on the nasal cavity by acoustic rhinometry and computed rhinomanometry. Twenty-nine children of both sexes with oral and/or mixed breathing, ranging in age from 7 to 10 years and with mixed dentition were selected. The children had uni- or bilateral posterior crossbite involving deciduous canines and the first permanent molars and were not being submitted to any otorhinolaryngologic or orthodontic treatment. All subjects were submitted to rhinologic exams and orthodontic documentation at three different times, i.e., before expansion and immediately and 90 days after expansion. There was no difference in the minimal cross-sectional area at the level of the valve and inferior nasal turbinate between the periods analyzed, but there was a statistically significant reduction in nasal resistance after expansion. On the basis of the present results, we may conclude that rapid maxillary expansion may lessen the nasal resistance. Although there was no difference in nasal geometry. Thus, this procedure may improve nasal respiration, but cannot be indicated for this purpose by itself.


Lipoprotein size is a main determinant for the rate of hydrolysis by exogenous LPL in human plasma.
LPL is a key player in plasma triglyceride metabolism. Consequently, LPL is regulated by several proteins during synthesis, folding, secretion, and transport to its site of action at the luminal side of capillaries, as well as during the catalytic reaction. Some proteins are well known, whereas others have been identified but are still not fully understood. We set out to study the effects of the natural variations in the plasma levels of all known LPL regulators on the activity of purified LPL added to samples of fasted plasma taken from 117 individuals. The enzymatic activity was measured at 25°C using isothermal titration calorimetry. This method allows quantification of the ability of an added fixed amount of exogenous LPL to hydrolyze triglyceride-rich lipoproteins in plasma samples by measuring the heat produced. Our results indicate that, under the conditions used, the normal variation in the endogenous levels of apolipoprotein C1, C2, and C3 or the levels of angiopoietin-like proteins 3, 4, and 8 in the fasted plasma samples had no significant effect on the recorded activity of the added LPL. Instead, the key determinant for the LPL activity was a lipid signature strongly correlated to the average size of the VLDL particles. The signature involved not only several lipoprotein and plasma lipid parameters but also apolipoprotein A5 levels. While the measurements cannot fully represent the action of LPL when attached to the capillary wall, our study provides knowledge on the interindividual variation of LPL lipolysis rates in human plasma.


Comparative Effectiveness of Infliximab and Adalimumab in Crohn's Disease and Ulcerative Colitis.
The availability of monoclonal antibodies to tumor necrosis factor α has revolutionized management of Crohn's disease (CD) and ulcerative colitis. However, limited data exist regarding comparative effectiveness of these agents to inform clinical practice. This study consisted of patients with CD or ulcerative colitis initiation either infliximab (IFX) or adalimumab (ADA) between 1998 and 2010. A validated likelihood of nonresponse classification score using frequency of narrative mentions of relevant symptoms in the electronic health record was applied to assess comparative effectiveness at 1 year. Inflammatory bowel disease-related surgery, hospitalization, and use of steroids were determined during this period. Our final cohort included 1060 new initiations of IFX (68% for CD) and 391 of ADA (79% for CD). In CD, the likelihood of nonresponse was higher in ADA than IFX (odds ratio, 1.62 and 95% CI, 1.21-2.17). Similar differences favoring efficacy of IFX were observed for the individual symptoms of diarrhea, pain, bleeding, and fatigue. However, there was no difference in inflammatory bowel disease-related surgery, hospitalizations, or prednisone use within 1 year after initiation of IFX or ADA in CD. There was no difference in narrative or codified outcomes between the 2 agents in ulcerative colitis. We identified a modestly higher likelihood of symptomatic nonresponse at 1 year for ADA compared with IFX in patients with CD. However, there were no differences in inflammatory bowel disease-related surgery or hospitalizations, suggesting these treatments are broadly comparable in effectiveness in routine clinical practice.


Impaired glutamine metabolism in NMDA receptor hypofunction induced by MK801.
Paradoxically, glutamate receptor antagonists have neurotoxic and psychotogenic properties in addition to their neuroprotective potential during excessive glutamate release. In the present study the non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK801 was used to examine glial-neuronal interactions in NMDA receptor hypofunction. Rats were given a subanesthetic dose of MK801 together with [1-13C]glucose and [1,2-13C]acetate, and brains were removed 20 min later. Analyses of extracts from cingulate, retrosplenial plus middle frontal cortices (CRFC) and temporal lobe were performed using HPLC and 13C and 1H nuclear magnetic resonance spectroscopy. Hypofunction of the NMDA receptor induced similar changes in both brain areas investigated; however, the changes were most pronounced in the temporal lobe. Generally, only labeling from [1-13C]glucose was affected by MK801. In CRFC and temporal lobe amounts of both labeled and unlabeled glutamine were increased, whereas those of aspartate were decreased. In the CRFC the decrease in labeling of aspartate was greater than the decrease in concentration, leading to decreased 13C enrichment. In temporal lobe, not in CRFC, increased concentrations of glutamate, GABA, succinate, glutathione and inositol were detected together with increased labeling of GABA and succinate from [1-13C]glucose. 13C Enrichment was decreased in glutamate and increased in succinate. The results point towards a disturbance in glutamate-glutamine cycling and thus interaction between neurons and glia, since labeling of glutamate and glutamine from glucose was affected differently.


Serum carnitine levels and carnitine esters of patients after kidney transplantation: role of immunosuppression.
Serum levels of carnitine and carnitine esters were measured in different groups of patients after cadaveric renal transplantation and compared with those of healthy subjects as well as azotemic and uremic patients. In all groups of patients serum levels of total carnitine (TC), free carnitine (FC), short-chain acylcarnitine (SCC), and long-chain acylcarnitine (LCC) increased with reduction of kidney function. However, cyclosporin- and prednisone-treated transplant patients with impaired kidney function displayed significantly higher TC, FC, and SCC compared with transplant patients under immunosuppression with azathioprine and prednisone. This group of cyclosporin-treated patients showed also significantly elevated serum cholesterol suggesting that carnitine deficiency is not responsible for the observed abnormalities in lipid metabolism after renal transplantation. Urinary excretion of TC, FC, SCC, and LCC decreased with reduction of kidney function without differences between cyclosporin- and azathioprine-treated patients. Single dose L-carnitine administration (10 mg/kg body weight IV) resulted in significantly higher TC, FC, SCC, and LCC values of azotemic patients with and without immunosuppression than in healthy subjects. Acylation of the administered carnitine was comparable in healthy controls and azotemic patients. Increased values of short-chain and long-chain acylcarnitine, therefore, seem to depend on the excretion of the diseased kidney(s). Our data demonstrate abnormalities in carnitine metabolism in patients with impaired kidney function. These alterations are further influenced by immunosuppressive drugs.


In Vitro Antifungal Activity of Ibrexafungerp (SCY-078) Against Contemporary Blood Isolates From Medically Relevant Species of Candida: A European Study.
Ibrexafungerp (SCY-078) is the newest oral and intravenous antifungal drug with broad activity, currently undergoing clinical trials for invasive candidiasis. The aim of this study was to assess the in vitro activity of ibrexafungerp and comparators against a collection of 434 European blood isolates of Candida. Ibrexafungerp, caspofungin, fluconazole, and micafungin minimum inhibitory concentrations (MICs) were collected from 12 European laboratories for 434 blood isolates, including 163 Candida albicans, 108 Candida parapsilosis, 60 Candida glabrata, 40 Candida tropicalis, 29 Candida krusei, 20 Candida orthopsilosis, 6 Candida guilliermondii, 2 Candida famata, 2 Candida lusitaniae, and 1 isolate each of Candida bracarensis, Candida catenulata, Candida dubliniensis, and Candida kefyr. MICs were determined by the EUCAST broth microdilution method, and isolates were classified according to recommended clinical breakpoints and epidemiological cutoffs. Additionally, 22 Candida auris from different clinical specimens were evaluated. Ibrexafungerp MICs ranged from 0.016 to ≥8 mg/L. The lowest ibrexafungerp MICs were observed for C. albicans (geometric MIC 0.062 mg/L, MIC range 0.016-0.5 mg/L) and the highest ibrexafungerp MICs were observed for C. tropicalis (geometric MIC 0.517 mg/L, MIC range 0.06-≥8 mg/L). Modal MICs/MIC50s (mg/L) against Candida spp. were 0.125/0.06 for C. albicans, 0.5/0.5 for C. parapsilosis, 0.25/0.25 for C. glabrata, 0.5/0.5 for C. tropicalis, 1/1 for C. krusei, 4/2 for C. orthopsilosis, and 0.5/0.5 for C. auris. Ibrexafungerp showed activity against fluconazole- and echinocandin-resistant isolates. If adopting wild-type upper limits, a non-wild-type phenotype for ibrexafungerp was only observed for 16/434 (3.7%) isolates: 11 (4.6%) C. parapsilosis, 4 (5%) C. glabrata, and 1 (2.5%) C. tropicalis. Ibrexafungerp showed a potent in vitro activity against Candida.


Predictability of spatial and non-spatial target properties improves perception in the pre-saccadic interval.
In a dual-task paradigm with a perceptual discrimination task and a concurrent saccade task, we examined participants' ability to make use of prior knowledge of a critical property of the perceptual target to improve discrimination. Previous research suggests that during a short time window before a saccade, covert attention is imperatively directed towards the saccade target location. Consequently, discrimination of perceptual targets at the saccade target location is better than at other locations. We asked whether the obligatory pre-saccadic attention shift prevents perceptual benefits arising for perceptual target stimuli with predictable as opposed to non-predictable properties. We compared conditions in which the color or location of the perceptual target was constant to conditions in which those properties varied randomly across trials. In addition to the expected improvements of perception at the saccade target location, we found perception to be better with constant than with random properties of the perceptual target. Thus, color or location information about an upcoming perceptual target facilitates perception even while spatial attention is shifted to the saccade target. The improvement occurred irrespective of the saccade target location, which suggests that the underlying mechanism is independent of the pre-saccadic attention shift, but alternative interpretations are discussed as well.


Mechanism of the decreased erythropoiesis in the water deprived rat.
Radioiron uptake by erythrocytes, metabolic rate, erythropoietin formation during hypoxia and erythroid responsiveness to exogenous erythropoietin were determined in both starved and water deprived rats. The feed intake showed a marked and progressive reduction during water deprivation. The metabolic rates of rats deprived of either food of water declined progressively showing a 40% reduction 5d after water deprivation or starvation began. At this time, the 24 h red blood cells 59Fe incorporation was 85% lower in both starved and dehydrated rats than in normal rats. Plasma erythropoietin levels in response to hypoxia were approximately 50% decreased in both starved and dehydrated rats. Both polycythaemic starved and polycythaemic water deprived rats injected with human urinary erythropoietin showed a 75% decrease in 59Fe incorporation into erythrocytes when compared to control rats. It is suggested that depression of erythropoiesis during water deprivation in the rat depends on a reduced sensitivity to erythropoietin, possibly associated with decreased production of the hormone. Since water deprived rats drastically reduce feed intake it is suggested that secondary starvation is the principal cause of the decreased erythropoiesis induced in the rat by water deprivation.


Organizational and market factors associated with Medicare dependence in inpatient rehabilitation hospitals.
Rehabilitation hospitals in the USA have been excluded from the Medicare Prospective Payment System (PPS) system since 1982, and have received cost-based reimbursement. However, the 1997 Balanced Budget Act mandated a PPS for inpatient rehabilitation, to be implemented by the end of 2002. This study assesses rehabilitation hospitals' dependency on Medicare. Findings show that not-for-profit hospitals, facilities with fewer services, facilities with lower staffing levels, and hospitals with lower operating expenses and profits, have a higher proportion of their inpatient revenue coming from Medicare. These facilities may be vulnerable to the new PPS payment system.


Factors influencing pulmonary surfactant protein A levels in cord blood, maternal blood and amniotic fluid.
The purpose of this study was to evaluate factors that influence pulmonary surfactant protein A (SP-A) levels in cord blood, maternal blood and amniotic fluid, as well as to establish the normal range of serum SP-A. Labor significantly influenced cord blood SP-A levels. The SP-A levels in maternal blood after delivery were correlated with, but higher than those before delivery. There was a correlation between SP-A levels in amniotic fluid and cord blood. Neonatal serum SP-A was not correlated with maternal SP-A levels. The normal range of SP-A in cord blood was 2.7-21.7 ng/ml following cesarean section without labor, 4.8-50.2 ng/ml after labor at gestational weeks 36-38, and 12.2-44.6 ng/ml at gestational weeks 39-41. SP-A levels in maternal blood before and after delivery and amniotic fluid were 6. 0-74.0, 9.6-73.6 and 403.4-24,540 ng/ml, respectively.


A case of focal nodular hyperplasia with a new characteristic finding on contrast-enhanced ultrasonography using Levovist.
We used contrast-enhanced ultrasound with Levovist, a microbubble contrast agent, to diagnose a case of hepatic focal nodular hyperplasia (FNH). A new characteristic finding of heartbeat-synchronized centrifugal enhancement was discovered. We call this enhancement pattern the "sonographic fireworks sign." It is expected to be useful for diagnosing FNH, especially when the lesions are small and it is difficult to depict a spoke-wheel pattern.


Black patients with COVID-19 had increased risk for 30-d mortality, PE, and other adverse outcomes vs. White patients.
Metra B, Summer R, Brooks SE, et al. Racial disparities in COVID-19 associated pulmonary embolism: a multicenter cohort study. Thromb Res. 2021;205:84-91. 34274560.


Pharmacokinetics of gentamicin in very low birth weight preterm infants.
Low birth weight preterm infants with suspected infection were administered gentamicin intramuscularly every 18 h (2.5 mg/kg) or 24 h (3.0 mg/kg). For both dosage regimens plasma gentamicin levels were monitored during a dosage interval on three separate occasions over a 10 day period. Both regimens gave satisfactory plasma concentrations and there was no important statistically significant difference between the two. The body clearance of gentamicin correlated with gestational age (r = 0.76, p less than 0.01). The results indicate either regimen may be useful in the clinical situation but from a practical standpoint administration every 24 h may be easier to comply with then every 18 h.


The efficacy of botulinum toxin for the treatment of trigeminal and postherpetic neuralgia: a systematic review with meta-analyses.
To evaluate the efficacy of a botulinum toxin type A (BoTN-A) in treating trigeminal neuralgia (TN) and postherpetic neuralgia (PHN). Three databases were searched: Medline, Web of Science, and Cochrane Library. The search was restricted to English-language randomized, placebo-controlled trials. Three review authors evaluated the cases for risk of bias. Six studies were eligible for inclusion. Pooled results showed a difference in post-treatment pain intensity of -3.009 (95% confidence interval -4.566 to -1.453; P < .001) in favor of BoTN-A compared with placebo in managing TN or PHN. Of the six studies, five had unclear risk of bias, and one showed high risk. Although the studies had unclear or high risk of bias, moderate evidence regarding the efficacy of BoTN-A in treating TN and PHN was found. BoTN-A might be an alternative treatment to those patients who are either unable to manage their pain medically or would like adjunct therapy.


[Effect of Inhibiting SIX1 Expression on Drug-resistance of Acute Myeloid Leukemia Cell Line HL-60/ADR Cells].
To establish HL-60 cells and adriamycin resistant HL-60 cells (H-60/ADR) in which the expression of homologous box gene 1 (SIX1) was inhibited, and investigate the effect of inhibiting the expression of SIX1 on the drug resistance. Lentivirus was used to transfect HL-60 and HL-60/ADR cells, and the cell lines stably inhibiting the expression of SIX1 were screened by puromycin. CCK-8 assay was used to detect the proliferation ability of cells in each group, apoptosis kit was used to detect the cell apoptosis, and real-time quantitative PCR was used to detect the expression level of drug-resistant related genes. HL-60 and HL-60/ADR stably transfected cell lines with down-regulation of SIX1 expression were successfully constructed. Compared with control group, the inhibition of SIX1 expression significantly inhibited the proliferation of HL-60 and HL-60/ADR cells (P <0.05), increased the apoptosis rate (P <0.05), and the sensitivity of cells to adriamycin increased after inhibition of SIX1 expression. Inhibition of SIX1 expression can improve cell sensitivity to adriamycin, and its role in reversing drug resistance may be related to the promotion of apoptosis gene expression.


Identification of 23-demethylacanthasterol in an asteroid, Acanthaster planci and its synthesis.
23-Demethylacanthasterol was identified in an asteroid, Acanthaster planci, by GC-MS analysis and direct comparison with the synthetic sample prepared from 23-demethylgorgosterol. The sterol composition of A. planci is also described.


Organophosphorus compounds biodegradation by novel bacterial isolates and their potential application in bioremediation of contaminated water.
Organophosphorus compounds (OPs), the major pesticides used worldwide, comprise an environmental hazard due to their harmful toxicity. Aimed to develop a bioreactor to remediate OPs contaminated wastewater, bacteria isolated from contaminated soils were identified and their ability to degrade OPs assessed, resulting in two main isolates, Sphingomonas sp. and Brevundimonas sp. Their OP degrading activities were characterized in terms of temperature, pH and substrates acceptance, resulting in high degradation rates at 60 °C, pH 10 and towards bulky OPs such as coroxon, coumaphos, and chlorpyrifos. Sphingomonas sp. cells were immobilized and 75.4% degradation of 0.15 mM chlorpyrifos was achieved after 21 days by immobilized cells in batch system, while this OP was completely degraded within 17 h when the biocatalyst is settled in a packed bed bioreactor, with a reusability of 8 cycles. These results suggest the potential application of this system in the bioremediation of contaminated wastewater.


Descemet stripping endothelial keratoplasty: effect of the surgical procedure on corneal optics.
To determine the changes in corneal optical performance after posterior lamellar corneal transplantation. Retrospective case series. The anterior segment in four eyes of four patients who underwent Descemet stripping endothelial keratoplasty (DSEK) with cataract extraction and intraocular lens (IOL) implantation were imaged with the Visante anterior segment optical coherence tomography [OCT] (Carl Zeiss Meditec, Dublin, California, USA). The curvature of the posterior surface of the donor graft was compared with that of the host cornea, and corneal thickness was measured. All eyes had a hyperopic refractive error after surgery. The posterior corneal curvature after surgery was more than that before surgery. Average preoperative keratometry was 43.4 diopters (D), and after surgery, it was 42.8 D using keratometry. However, when the postsurgical corneal power was calculated using the Gaussian optics method, the average value was 40.8 D. The addition of a donor corneal graft to the posterior surface of decompensated corneas may lessen the effective optical power of the cornea and may have implications for IOL power calculations in these eyes.


The extracellular death factor (EDF) protects Escherichia coli by scavenging hydroxyl radicals induced by bactericidal antibiotics.
The newly discovered extracellular death factor (EDF) is a pentapeptide with the sequence NNWNN in Escherichia coli. It was reported that it participated in the cell death process mediated by toxin-antitoxin system mazEF. Reactive oxygen species (ROS) are recently considered as common factors for bactericidal antibiotics-mediated cell death. Previous study indicated that EDF could scavenge hydroxyl radicals and might act as a signal molecule with dual effects, "death" and "survival". But the structure-activity relationship of EDF and the effects of EDF on the activity of antibiotics remain unclear. In the present study, our results indicated that tryptophan could be the key residue to the hydroxyl radicals-scavenging activity of EDF, and EDF could protect Escherichia coli from killing by bactericidal antibiotics, but not by DNA-damaging or bacteriostatic antibiotics. Our results could provide novel evidence to understand the role of EDF in drug-resistance.


[The role of telocytes in myocardial remodeling and the development of cardiovascular complications in patients with chronic heart failure after coronary artery bypass grafting].
Аim of the study. A study of the role of cardiac telocytes (Tc) in myocardial remodeling and the effect of changes in their morphology on the clinical outcome of coronary bypass (CABG) in patients with chronic heart failure (CHF). The perioperative period was analyzed in 83 patients (43 men and 40 women) with CHF I, IIA and IIB, NYHA functional class I-III undergoing CABG. Myocardial biopsy of the right atrial appendage (RAA), obtained during the CABG, was studied using histological, immunohistochemical (expression of CD-34/vimentin, MMP-2, TIMP-1, caspase-3, bcl-2) and morphometric methods. In the analysis of the myocardium of RAA of a group of patients with cardiovascular complications (CVC) after CABG, a significant decrease in the diameter of cardiomyocytes (CMCs) of their nuclei, bulk density (BD) of CMC, a significant decrease in BD Tc, trophic index; an increase in CMC with signs of apoptosis, BD of stroma, a violation of the ratio of MMP-2/TIMP-1. The intercellular network Tc with secreted regulatory factors and microvesicles is an important structural and functional link of the myocardium. The decrease in BD Tc has a significant effect on the remodeling and regenerative potential of the myocardium, leading to the development of CVC in patients with CHF.


Influence of physical factors on the development of gastroesophageal reflux in man: analysis by means of an experimental mechanical model.
In this paper, the isolated influence of physical factors on the development of gastroesophageal reflux in man is analyzed by means of a mechanical model. We evaluate the influence of gravity, intraabdominal pressure, intrathoracic pressure, transmission of intraabdominal pressure to the high pressure zone, filling volume of the stomach, and the existence of a high pressure zone on the appearance of gastroesophageal reflux. PH-metry is used to record the episodes of reflux in the model. We wish to demonstrate the importance of gravity and intraabdominal pressure in the production of reflux. Intrathoracic pressure acts as an antireflux mechanism only when associated with a zone of high pressure. The intraabdominal pressure may be transmitted to the high pressure zone as a purely mechanical effect and the greater gastric filling volume may, when associated with the intraabdominal pressure, facilitate reflux without physical changes in the high pressure zone (the equivalent of a reduction in the length of the lower esophageal sphincter in man). We conclude that this model may help to explain the influence of the physical factors mentioned above on the development of gastroesophageal reflux in man.


Evaluation of PUVA, topical corticosteroids and the combination of both in the treatment of psoriasis.
A total of 90 patients with psoriasis were treated either by photochemotherapy (PUVA) or by topical steroids under occlusion (TOC). In a third group of patients a combination of both regimens was applied. All patients received approximately 15 treatments of either type. During therapy, biopsies were taken in all treatment groups and the epidermal thickness was monitored planimetrically. After clearing of the skin the patients were left without treatment and the time of onset of new lesions was recorded. While TOC produced rapid clearing of the skin this was followed by early relapses (in 50% of the patients after 3 weeks). PUVA reduced psoriatic hyperplasia more slowly; however, 50% of the patients remained free of psoriasis for approximately 10 weeks after clearance. Psoriasis treated with steroids under occlusion together with PUVA showed a significantly faster return of the skin to normal as compared to PUVA. As in PUVA-treated patients relapses occurred after approximately 12 weeks is this group. Therefore the combination of both treatments appears to be advantageous for rapid skin clearance and comparatively long remissions.


Early initial video-electro-encephalography combined with variant location predict prognosis of KCNQ2-related disorder.
The clinical features of KCNQ2-related disorders range from benign familial neonatal seizures 1 to early infantile epileptic encephalopathy 7. The genotype-phenotypic association is difficult to establish. To explore potential factors in neonatal period that can predict the prognosis of neonates with KCNQ2-related disorder. Infants with KCNQ2-related disorder were retrospectively enrolled in our study in Children's Hospital of Fudan University in China from Jan 2015 to Mar 2020. All infants were older than age of 12 months at time of follow-up, and assessed by Bayley Scales of Infant and Toddler Development-Third Edition (BSID-III) or Wechsler preschool and primary scale of intelligence-fourth edition (WPPSI-IV), then divided into three groups based on scores of BSID-III or WPPSI-IV: normal group, mild impairment group, encephalopathy group. We collected demographic variables, clinical characteristics, neuroimaging data. Considered variables include gender, gestational age, birth weight, age of the initial seizures, early interictal VEEG, variant location, delivery type. Variables predicting prognosis were identified using multivariate ordinal logistic regression analysis. A total of 52 infants were selected in this study. Early interictal video-electro-encephalography (VEEG) (β = 2.77, 1.20 to 4.34, P = 0.001), and variant location (β = 2.77, 0.03 to 5.5, P = 0.048) were independent risk factors for prognosis. The worse the early interictal VEEG, the worse the prognosis. Patients with variants located in the pore-lining domain or S4 segment are more likely to have a poor prognosis. The integration of early initial VEEG and variant location can predict prognosis. An individual whose KCNQ2 variant located in voltage sensor, the pore domain, with worse early initial VEEG background, often had an adverse outcome.


Polymerization degree of chitosan affects structural and compositional changes in the cell walls, membrane lipids, and proteins in the leaves of Ipomoea purpurea: An FT-IR spectroscopy study.
This research aimed to investigate the polymerization degree (DP) -dependent effects of chitosan treatments on structural and compositional changes in certain cell wall polysaccharides (mainly lignin), membrane lipids, and proteins of in vitro-propagated Ipomoea purpurea leaves through FT-IR spectroscopy. The chitosan oligomer mixture (DP 2-15; CHI-OM) and chitosan polymer (DP 70; CHI-P) applied at 5, 10, and 20 mg L-1 concentrations induced different patterns of biomolecular changes in I. purpurea leaves. The chitosan variants enhanced the destabilization of cell membrane structures. CHI-P treatments increased the lipid structure and protein content of the membranes more than CHI-OM treatments. CHI-OM treatment was found to have the ability to induce the formation of β-sheet structures with a low number of strands, whereas CHI-P treatment was found to have the ability to trigger the formation of more extended α-helix structures in the secondary structure of proteins. CHI-P treatments increased lignin synthesis more than CHI-OM treatments. However, CHI-OM at 10 mg L-1 concentration was more effective than CHI-P treatments in the induction of cell wall polysaccharide synthesis. These findings suggest that the polymerization degree of chitosan plays a role in changing structures and compositions of the biomolecules present in the leaves of I. purpurea.


Detection of cervical intraepithelial neoplasia in women with atypical squamous or glandular cells of undetermined significance cytology: a prospective study.
(1) To assess the prevalence of histologically confirmed cervical intraepithelial neoplasia in patients with cervical smears diagnosed as atypical squamous or glandular cells of undetermined significance. (2) To evaluate the role of colposcopy and the presence of human papillomavirus in detecting underlying cervical intraepithelial neoplasia. In this prospective cohort, 148 women with atypical squamous or glandular cells of undetermined significance were evaluated by colposcopy, histological sampling, and human papillomavirus deoxyribonucleic acid testing. Histological diagnosis of >/= cervical intraepithelial neoplasia II was found in 10/148 women. Women with a histological >/= cervical intraepithelial neoplasia II had a higher prevalence of >/= two abnormal quadrants (90% vs. 38%</= cervical intraepithelial neoplasia I, p = 0.002) and of high/intermediate-risk human papillomavirus (90% vs. 42% </= cervical intraepithelial neoplasia I, p = 0.005). Our study shows that premalignant lesions of the cervix were present in 7% of the patients with a cytological diagnosis of atypical squamous or glandular cells of undetermined significance. Colposcopy and human papillomavirus deoxyribonucleic acid testing are both important parameters in detecting >/= cervical intraepithelial neoplasia II.


Impaired aldehyde dehydrogenase 1 subfamily member 2A-dependent retinoic acid signaling is related with a mesenchymal-like phenotype and an unfavorable prognosis of head and neck squamous cell carcinoma.
An inverse correlation between expression of the aldehyde dehydrogenase 1 subfamily A2 (ALDH1A2) and gene promoter methylation has been identified as a common feature of oropharyngeal squamous cell carcinoma (OPSCC). Moreover, low ALDH1A2 expression was associated with an unfavorable prognosis of OPSCC patients, however the causal link between reduced ALDH1A2 function and treatment failure has not been addressed so far. Serial sections from tissue microarrays of patients with primary OPSCC (n = 101) were stained by immunohistochemistry for key regulators of retinoic acid (RA) signaling, including ALDH1A2. Survival with respect to these regulators was investigated by univariate Kaplan-Meier analysis and multivariate Cox regression proportional hazard models. The impact of ALDH1A2-RAR signaling on tumor-relevant processes was addressed in established tumor cell lines and in an orthotopic mouse xenograft model. Immunohistochemical analysis showed an improved prognosis of ALDH1A2(high) OPSCC only in the presence of CRABP2, an intracellular RA transporter. Moreover, an ALDH1A2(high)CRABP2(high) staining pattern served as an independent predictor for progression-free (HR: 0.395, p = 0.007) and overall survival (HR: 0.303, p = 0.002), suggesting a critical impact of RA metabolism and signaling on clinical outcome. Functionally, ALDH1A2 expression and activity in tumor cell lines were related to RA levels. While administration of retinoids inhibited clonogenic growth and proliferation, the pharmacological inhibition of ALDH1A2-RAR signaling resulted in loss of cell-cell adhesion and a mesenchymal-like phenotype. Xenograft tumors derived from FaDu cells with stable silencing of ALDH1A2 and primary tumors from OPSCC patients with low ALDH1A2 expression exhibited a mesenchymal-like phenotype characterized by vimentin expression. This study has unraveled a critical role of ALDH1A2-RAR signaling in the pathogenesis of head and neck cancer and our data implicate that patients with ALDH1A2(low) tumors might benefit from adjuvant treatment with retinoids.


Nonablative phototherapy for acne vulgaris using the KTP 532 nm laser.
Acne vulgaris is an acute inflammatory disease of the pilosebaceous units. The bastion of treatment for acne vulgaris has been the use of topical and systemic therapies. Despite many modalities available for treatment, there exists an imperative need for effective noninvasive treatments that reduce the risks of medication side effects. To study the safety and efficacy of the potassium titanyl phosphate (KTP) 532 nm pulsed laser for the treatment of acne vulgaris. Twenty-six subjects, clinically evaluated with moderate facial acne, were enrolled in this single-center prospective trial. The entire facial area for each subject was divided in half and randomly designated as either a treatment or a control side. Each subject was treated with four laser exposures using a KTP 532 nm laser with continuous contact cooling. The results were assessed at 1 and 4 weeks post-final treatment. Primary outcome measures were Michaëlsson acne severity score and adverse treatment effects. Secondary outcome measures included subjective evaluations from the investigator and patients assessing their overall percent satisfaction. Primary outcome analysis in the Michaëlsson acne severity score demonstrated a mean 34.9% (p = .011) and 20.7% (p = .25) reduction at the 1-week and 4-week post-final treatments, respectively. Subjective investigator evaluations of overall percent satisfaction indicated that all patients demonstrated a minimum 50% overall satisfaction in treatment outcomes at the 4-week follow-up period. No side effects were encountered. Use of the KTP 532 nm laser for the treatment and management of acne vulgaris is both safe and effective, with positive results enduring up to 4 weeks post-treatment.


Is the pencil point spinal needle a better choice in younger patients? A comparison of 24G Sprotte with 27G Quincke needles in an unselected group of general surgical patients below 46 years of age.
Reports have indicated that there are less postoperative complaints after the use of pencil pointed spinal needles. We compared a 24G Sprotte needle with a 27G Quincke needle in a randomised study of 200 healthy patients (49% females), aged 15-46 years. Four patients (2%) reported postdural puncture headache, three with the 24G Sprotte needle and one with the 27G Quincke needle. Thirteen patients (7%) suffered with nonspecific headache, with no significant difference between the two groups. Of the 57 (29%) who reported backpain, a significantly higher proportion had received spinal anaesthesia with the Sprotte needle (OR = 2.06). There was a significantly higher incidence of insufficient blocks after dural puncture with the Sprotte needle. Ease of needle insertion and number of puncture attempts was the same for both needle types.


Natural killer activity in the experimental privational rickets.
To study the 'in vivo' importance of vitamin D on the natural killer (NK) activity, rats were submitted to privational rickets induced by a diet deficient in vitamin D and phosphorus (D-P-). Thirty days after the beginning of treatment the animals showed low body weight, changes in the bone development, and decreased levels of 25-hydroxyvitamin D(3) (25-OH D(3)). NK activity, evaluated using a cytotoxicity assay against 51Cr-labeled Yac.1 target cells, was not modified by the rickets-inducing treatment during the first 30 days. Following a long-term treatment (60 days) the rachitic rats (D-P-) exhibited higher NK activity than control animals (D+P+) (P<0.05). On the other hand, D-P+ animals showed higher cytotoxic activity than D-P- and D+P+ groups. Feed replacement to the rachitic rats by a complete diet (D-P-/D+P+) led to a partial recuperation of growth, bone development, and 25-OH D(3) serum levels. The NK activity was also influenced by vitamin D intake, decreasing after treatment.


TRPC6, a therapeutic target for pulmonary hypertension.
Idiopathic pulmonary arterial hypertension (PAH) is a fatal and progressive disease. Sustained vasoconstriction due to pulmonary arterial smooth muscle cell (PASMC) contraction and concentric arterial remodeling due partially to PASMC proliferation are the major causes for increased pulmonary vascular resistance and increased pulmonary arterial pressure in patients with precapillary pulmonary hypertension (PH) including PAH and PH due to respiratory diseases or hypoxemia. We and others observed upregulation of TRPC6 channels in PASMCs from patients with PAH. A rise in cytosolic Ca2+ concentration ([Ca2+]cyt) in PASMC triggers PASMC contraction and vasoconstriction, while Ca2+-dependent activation of PI3K/AKT/mTOR pathway is a pivotal signaling cascade for cell proliferation and gene expression. Despite evidence supporting a pathological role of TRPC6, no selective and orally bioavailable TRPC6 antagonist has yet been developed and tested for treatment of PAH or PH. In this study, we sought to investigate whether block of receptor-operated Ca2+ channels using a nonselective blocker of cation channels, 2-aminoethyl diphenylborinate (2-APB, administered intraperitoneally) and a selective blocker of TRPC6, BI-749327 (administered orally) can reverse established PH in mice. The results from the study show that intrapulmonary application of 2-APB (40 µM) or BI-749327 (3-10 µM) significantly and reversibly inhibited acute alveolar hypoxia-induced pulmonary vasoconstriction. Intraperitoneal injection of 2-APB (1 mg/kg per day) significantly attenuated the development of PH and partially reversed established PH in mice. Oral gavage of BI-749327 (30 mg/kg, every day, for 2 wk) reversed established PH by ∼50% via regression of pulmonary vascular remodeling. Furthermore, 2-APB and BI-749327 both significantly inhibited PDGF- and serum-mediated phosphorylation of AKT and mTOR in PASMC. In summary, the receptor-operated and mechanosensitive TRPC6 channel is a good target for developing novel treatment for PAH/PH. BI-749327, a selective TRPC6 blocker, is potentially a novel and effective drug for treating PAH and PH due to respiratory diseases or hypoxemia.


Renewal of oligodendrocyte lineage reverses dysmyelination and CNS neurodegeneration through corrected N-acetylaspartate metabolism.
Myelinating oligodendrocytes are essential for neuronal communication and homeostasis of the central nervous system (CNS). One of the most abundant molecules in the mammalian CNS is N-acetylaspartate (NAA), which is catabolized into L-aspartate and acetate by the enzyme aspartoacylase (ASPA) in oligodendrocytes. The resulting acetate moiety is thought to contribute to myelin lipid synthesis. In addition, affected NAA metabolism has been implicated in several neurological disorders, including leukodystrophies and demyelinating diseases such as multiple sclerosis. Genetic disruption of ASPA function causes Canavan disease, which is hallmarked by increased NAA levels, myelin and neuronal loss, large vacuole formation in the CNS, and early death in childhood. Although NAA's direct role in the CNS is inconclusive, in peripheral adipose tissue, NAA-derived acetate has been found to modify histones, a mechanism known to be involved in epigenetic regulation of cell differentiation. We hypothesize that a lack of cellular differentiation in the brain contributes to the disruption of myelination and neurodegeneration in diseases with altered NAA metabolism, such as Canavan disease. Our study demonstrates that loss of functional Aspa in mice disrupts myelination and shifts the transcriptional expression of neuronal and oligodendrocyte markers towards less differentiated stages in a spatiotemporal manner. Upon re-expression of ASPA, these oligodendrocyte and neuronal lineage markers are either improved or normalized, suggesting that NAA breakdown by Aspa plays an essential role in the maturation of neurons and oligodendrocytes. Also, this effect of ASPA re-expression is blunted in old mice, potentially due to limited ability of neuronal, rather than oligodendrocyte, recovery.


Feeding Date-Palm Leaves Ensiled with Fibrolytic Enzymes or Multi-Species Probiotics to Farafra Ewes: Intake, Digestibility, Ruminal Fermentation, Blood Chemistry, Milk Production and Milk Fatty Acid Profile.
The present experiment evaluated the feeding of date palm leaves (DPL) ensiled with fibrolytic enzymes (ENZ) or multi-species probiotics (MSP) on nutrient utilization and lactational performance of ewes. Fifty multiparous lactating Farafra ewes were used in a completely randomized design for 90 d. The treatments consisted of the control diet with a concentrate feed mixture and date palm leaves (at 60:40, DM basis) ensiled without additive (control) or DPL ensiled with ENZ or MSP replacing control DPL at 50 or 100%. Both ENZ and MSP increased (p < 0.01) DPL and total intakes, digestibility of all nutrients, concentrations of ammonia, total volatile fatty acids, acetate and propionate in the rumen. Increased milk production, concentrations of fat, lactose and energy in milk, and feed efficiency were observed with MSP and ENZ compared to the control treatment. Moreover, ENZ and MSP increased (p < 0.05) the concentrations of total n3, n6 fatty acids, polyunsaturated fatty acids and conjugated linoleic acids and decreased (p < 0.001) the atherogenicity. The differences between ENZ and MSP and between the low and high replacement levels were minor for all measured parameters. Ensiling of DPL with MSP or fibrolytic enzymes is recommended to improve feed efficiency and improve lactational performance of ewes.


SARS-CoV-2 envelope protein triggers depression-like behaviors and dysosmia via TLR2-mediated neuroinflammation in mice.
Depression and dysosmia have been regarded as primary neurological symptoms in COVID-19 patients, the mechanism of which remains unclear. Current studies have demonstrated that the SARS-CoV-2 envelope (E) protein is a pro-inflammatory factor sensed by Toll-like receptor 2 (TLR2), suggesting the pathological feature of E protein is independent of viral infection. In this study, we aim to ascertain the role of E protein in depression, dysosmia and associated neuroinflammation in the central nervous system (CNS). Depression-like behaviors and olfactory function were observed in both female and male mice receiving intracisternal injection of E protein. Immunohistochemistry was applied in conjunction with RT-PCR to evaluate glial activation, blood-brain barrier status and mediators synthesis in the cortex, hippocampus and olfactory bulb. TLR2 was pharmacologically blocked to determine its role in E protein-related depression-like behaviors and dysosmia in mice. Intracisternal injection of E protein evoked depression-like behaviors and dysosmia in both female and male mice. Immunohistochemistry suggested that the E protein upregulated IBA1 and GFAP in the cortex, hippocampus and olfactory bulb, while ZO-1 was downregulated. Moreover, IL-1β, TNF-α, IL-6, CCL2, MMP2 and CSF1 were upregulated in both cortex and hippocampus, whereas IL-1β, IL-6 and CCL2 were upregulated in the olfactory bulb. Furtherly, inhibiting microglia, rather than astrocytes, alleviated depression-like behaviors and dysosmia induced by E protein. Finally, RT-PCR and immunohistochemistry suggested that TLR2 was upregulated in the cortex, hippocampus and olfactory bulb, the blocking of which mitigated depression-like behaviors and dysosmia induced by E protein. Our study demonstrates that envelope protein could directly induce depression-like behaviors, dysosmia, and obvious neuroinflammation in CNS. TLR2 mediated depression-like behaviors and dysosmia induced by envelope protein, which could serve as a promising therapeutic target for neurological manifestation in COVID-19 patients.


A case of hypertrophic protein losing gastropathy.
A 3 1/2 year old boy presented with a history of vomiting and generalized oedema. Biochemically proven protein losing enteropathy was associated with huge gastric fundal rugae on barium examination. Gastroscopy confirmed the barium findings but biopsy material demonstrated normal mucosa. The condition regressed on a high protein diet. Cytomegalovirus (CMV) was found in the urine after the illness had subsided.


[Clinical suggestions on the use of calcium hydroxide in therapy of pulp necrosis].
The Aa. discuss the Bernard's method for therapy of pulpar necrosis, based on using of calcium oxide, know as "ocalexic method". They report clinical procedure and results of their experience.


Endothelium-dependent inhibition of sympathetic vasoconstriction by frusemide administration to rats.
1. Segments of the tail artery of the rat were cannulated at both ends and mounted in an organ bath filled with Krebs solution. 2. Using an extracorporeal circuit, blood was pumped at a constant 2 ml/min from the carotid artery of anaesthetized rats to perfuse the segment of tail artery and returned to the donor rat via the jugular vein. 3. Peri-arterial electrical stimulation of the ex vivo blood perfused tail artery at 5 Hz produced vasoconstriction and an increase in perfusion pressure. 4. The intravenous administration of frusemide 5 mg/kg to the donor rat resulted in an inhibition of the vasoconstrictor responses of the perfused tail artery segment. Diuresis-induced losses of volume and frusemide were prevented by a urinary bladder-venous shunt. 5. Removal of the endothelium from the tail artery segment, by perfusion with dry gas for 4 min, prevented the vasoconstrictor-inhibitory effect of frusemide administration. Removal of the endothelium was confirmed histologically and by the absence of a vasodilator response to acetylcholine. 6. On the basis of these and previous results it is concluded that parenteral frusemide administration releases an unidentified but non-prostanoid hormone from the kidney which produces an endothelium-dependent inhibition of sympathetic vasoconstriction.


I131-rose bengal scanning in the detection of cholecystocolic fistula. New use of an established procedure.
A case of cholecystocolic fistula diagnosed by I131-Rose bengal scanning and later confirmed at the time of laparotomy is reported. The fistula was not seen by conventional contrast radiographic studies. It is suggested that I131-Rose bengal studies be considered in the study of patients suspected of having cholecystenteric fistulas.


Genetic and maternal effect influences on viability of common frog tadpoles under different environmental conditions.
The influence of environmental stress on the expression of genetic and maternal effects on the viability traits has seldom been assessed in wild vertebrates. We have estimated genetic and maternal effects on the viability (viz probability of survival, probability of being deformed, and body size and shape) of common frog, Rana temporaria, tadpoles under stressful (low pH) and nonstressful (neutral pH) environmental conditions. A Bayesian analysis using generalized linear mixed models was applied to data from a factorial laboratory experiment. The expression of additive genetic variance was independent of pH treatments, and all traits were significantly heritable (survival: h2 approximately 0.08; deformities: h2 approximately 0.26; body size: h2 approximately 0.12; body shape: h2 approximately 0.14). Likewise, nonadditive genetic contributions to variation in all traits were significant, independent of pH treatments and typically of magnitude similar to the additive genetic effects. Maternal effects were large for all traits, especially for viability itself, and their expression was partly dependent on the environment. In the case of body size, the maternal effects were mediated largely through egg size. In general, the results give little evidence for the conjecture that environmental stress created by low pH would impact strongly on the genetic architecture of fitness-related traits in frogs, and hamper adaptation to stress caused by acidification. The low heritabilities and high dominance contributions conform to the pattern typical for traits subject to relatively strong directional selection.