From bad behaviour to bad biology: pitfalls and promises in the management of resistant hypertension.
Control rates for hypertension have dramatically improved during the past 2 decades-especially in Canada. However, hypertension remains one of the top risk factors for premature death globally. Furthermore, one-third of Canadians with hypertension have not obtained adequate blood pressure control. Most of these patients have resistant hypertension with uncontrolled blood pressure despite therapy. The etiology of resistant hypertension is multifactorial but includes both behavioural and biological factors. Among behavioural factors, nonadherence on the part of patients and especially clinical inertia on the part of health care professionals are contributing causes. An understanding of the root causes underlying the failure to control an individual's blood pressure is central to optimal subsequent management. Further advances in blood pressure control rates in this group of patients will depend on improvements in health care delivery systems and the further development of innovative therapies. Drugs combining multiple antihypertensive agents in a single pill and the development of new technologies to lower blood pressure, primarily by disruption of the sympathetic nervous system, have the potential to be useful strategies in this effort.


Genetic challenges of flavor improvement in tomato.
In many instances, the intensive breeding of crops over the past half century with a focus on yield has indirectly led to reductions in flavor and nutrient content. Largely, this deterioration of quality relates directly to the genetic and biochemical complexity of such traits. Here, we describe challenges associated with quality improvement, emphasizing tomato fruit flavor. Flavor improvement is particularly problematic because of the difficulty of assessing the phenotype as well as a lack of fundamental knowledge about the chemicals driving consumer preferences, the pathways for their synthesis, and the genes regulating the output of these pathways. Recent breakthroughs from a systematic analysis of these factors and the availability of a tomato genome sequence have led to significant progress in our understanding of flavor. However, the need to deliver improved flavor in the context of high yield and long postharvest shelf life still present major challenges.


Histological prognosticators in neoadjuvant naive oesophageal cancer patients.
Prognosis of oesophageal cancer is primarily based upon the TNM stage of the disease. However, even in those with similar TNM staging, survival can be varied. Additional histopathological factors including venous invasion (VI), lymphatic invasion (LI) and perineural invasion (PNI) have been identified as prognostic markers yet are not part of TNM classification. The aim of this study is to determine the prognostic importance of these factors and overall survival in patients with oesophageal or junctional cancer who underwent transthoracic oesophagectomy as the unimodality treatment. Data from patients who underwent transthoracic oesophagectomy for adenocarcinoma without neoadjuvant treatment were reviewed. Patients were treated with radical resection, with a curative intent using a transthoracic Ivor Lewis or three staged McKeown approach. A total of 172 patients were included. Survival was poorer when VI, LI and PNI were present (p<0.001), with the estimated survival being significantly worse (p<0.001) when patients were stratified according to the number of factors present. Univariable analysis of factors revealed VI, LI and PNI were all associated with survival. Presence of LI was independently predictive of incorrect staging/upstaging in multivariable logistic regression analysis (OR 12.9 95% CI 3.6-46.6, p<0.001). Histological factors of VI, LI and PNI are markers of aggressive disease and may have a role in prognostication and decision-making prior to treatment. The presence of LI as an independent marker of upstaging could be a potential indication for the use of neoadjuvant treatment in patients with early clinical disease.


Methylmethacrylate hypersensitivity in a patient with cemented endoprosthesis. A case report.
A case of methylmethacrylate monomer hypersensitivity in a 76-year-old patient with a cemented endoprosthesis is reported. The accuracy of the method for preoperative testing of a patient's sensitivity is discussed.


When can we reconstruct the ancestral state? Beyond Brownian motion.
Reconstructing the ancestral state of a group of species helps answer many important questions in evolutionary biology. Therefore, it is crucial to understand when we can estimate the ancestral state accurately. Previous works provide a necessary and sufficient condition, called the big bang condition, for the existence of an accurate reconstruction method under discrete trait evolution models and the Brownian motion model. In this paper, we extend this result to a wide range of continuous trait evolution models. In particular, we consider a general setting where continuous traits evolve along the tree according to stochastic processes that satisfy some regularity conditions. We verify these conditions for popular continuous trait evolution models including Ornstein-Uhlenbeck, reflected Brownian Motion, bounded Brownian Motion, and Cox-Ingersoll-Ross.


Differential effects of "Advanced glycation endproducts" and beta-amyloid peptide on glucose utilization and ATP levels in the neuronal cell line SH-SY5Y.
Beta-amyloid peptide (Abeta) and "Advanced glycation endproducts" (AGEs) are components of the senile plaques in Alzheimer's disease patients. It has been proposed that both AGEs and Abeta exert many of their effects, which include the upregulation of pro-inflammatory cytokines, through RAGE ("receptor for advanced glycation endproducts"). To investigate whether Abeta and AGEs cause similar or identical effects on cell survival and energy metabolism, we have compared the effects of a model-AGE and Abeta on cell viability, ATP level, glucose consumption and lactate production in the neuroblastoma cell line SH-SY5Y. The results show that AGEs and Abeta increase glucose consumption and decrease ATP levels in a dose dependent manner. Furthermore, both compounds decrease mitochondrial activity measured by the MTT assay. However, only AGEs decrease the number of cells and significantly increase lactate production. These data indicate that both AGEs and Abeta can cause differential disturbances in neuronal metabolism, which may contribute to the pathophysiological findings in Alzheimer's disease. However, their signalling pathways are apparently quite distinct, a fact which should stimulate a more detailed investigation in this field, e.g. for the purpose of a rational design of potential "neuroprotective" RAGE antagonists.


Cholinergic enhancement augments magnitude and specificity of visual perceptual learning in healthy humans.
Learning through experience underlies the ability to adapt to novel tasks and unfamiliar environments. However, learning must be regulated so that relevant aspects of the environment are selectively encoded. Acetylcholine (ACh) has been suggested to regulate learning by enhancing the responses of sensory cortical neurons to behaviorally relevant stimuli. In this study, we increased synaptic levels of ACh in the brains of healthy human subjects with the cholinesterase inhibitor donepezil (trade name: Aricept) and measured the effects of this cholinergic enhancement on visual perceptual learning. Each subject completed two 5 day courses of training on a motion direction discrimination task, once while ingesting 5 mg of donepezil before every training session and once while placebo was administered. We found that cholinergic enhancement augmented perceptual learning for stimuli having the same direction of motion and visual field location used during training. In addition, perceptual learning with donepezil was more selective to the trained direction of motion and visual field location. These results, combined with previous studies demonstrating an increase in neuronal selectivity following cholinergic enhancement, suggest a possible mechanism by which ACh augments neural plasticity by directing activity to populations of neurons that encode behaviorally relevant stimulus features.


[Neuroendocrine differentiation in prostate cancer: an unrecognized and therapy resistant phenotype].
Neuroendocrine (NE) differentiation frequently occurs in common prostatic malignancies but usually escapes pathological and clinical detection. The present review focuses on biological properties of NE tumor cells making them resistant to androgen deprivation and radiation therapy. Recent data have shown that NE prostate cancer cells (as defined by the most commonly used endocrine marker chromogranin A) are arrested in the G0-phase of the cell cycle and do not undergo apoptosis. This particular phenotype consistently lacks the nuclear androgen receptor in both benign and malignant conditions but produces a series of hormonal growth factors exerting mitogenic stimuli on adjacent, exocrine tumor cells. Neoplastic NE cells devoid of the nuclear androgen receptor constitute an androgen-insensitive cell population in prostate cancer. The absence of proliferative and apoptotic activity makes NE tumor cells particularly resistant towards cytotoxic drugs and radiation therapy. Pathological and clinical detection of NE features is recommended for all prostate cancer patients for whom radiation therapy and androgen deprivation is being considered.


Quantification of myocardial fibrosis using noninvasive T2-mapping magnetic resonance imaging: Preclinical models of aging and pressure overload.
Noninvasive imaging of cardiac fibrosis is important for early diagnosis and intervention in chronic heart diseases. Here, we investigated whether noninvasive, contrast agent-free MRI T2 -mapping can quantify myocardial fibrosis in preclinical models of aging and pressure overload. Myocardial fibrosis and remodeling were analyzed in two animal models: (i) aging (15-month-old male CF-1 mice vs. young 6- to 8-week-old mice), and (ii) pressure overload (PO; by transverse aortic constriction in 4- to 5-month-old male C57BL/6 mice vs. sham-operated for 14 days). In vivo T2 -mapping was performed by acquiring data during the isovolumic and early diastolic phases, with a modified respiratory and ECG-triggered multiecho TurboRARE sequence on a 7-T MRI. Cine MRI provided cardiac morphology and function. A quantitative segmentation method was developed to analyze the in vivo T2 -maps of hearts at midventricle, apex, and basal regions. The cardiac fibrosis area was analyzed ex vivo by picro sirius red (PSR) staining. Both aged and pressure-overloaded hearts developed significant myocardial contractile dysfunction, cardiac hypertrophy, and interstitial fibrosis. The aged mice had two phenotypes, fibrotic and mild-fibrotic. Notably, the aged fibrotic subgroup and the PO mice showed a marked decrease in T2 relaxation times (25.3 ± 0.6 in aged vs. 29.9 ± 0.7 ms in young mice, p = 0.002; and 24.3 ± 1.7 in PO vs. 28.7 ± 0.7 ms in shams, p = 0.05). However, no significant difference in T2 was detected between the aged mild-fibrotic subgroup and the young mice. Accordingly, an inverse correlation between myocardial fibrosis percentage (FP) and T2 relaxation time was derived (R2 = 0.98): T2 (ms) = 30.45 - 1.05 × FP. Thus, these results demonstrate a statistical agreement between T2 -map-quantified fibrosis and PSR staining in two different clinically relevant animal models. In conclusion, T2 -mapping MRI is a promising noninvasive contrast agent-free quantitative technique to characterize myocardial fibrosis.


Hashish smoke interfers with Sidman avoidance in mice.
Hashish smoke has been proved to be active in the Sidman avoidance. Its activity is similar to that of hallucinogens.


Synaptic regulation of hypothalamic neurons containing ACTH by substance P.
Synaptic connections between substance P (SP)- and ACTH-containing neurons in the rat arcuate nucleus were studied by a technique of combined pre- and postembedding immunohistochemical staining. The results showed that immunoreactive (ir) SP fiber terminals form synapses with ir ACTH cell bodies. This strongly suggests that the activity of ACTH neurons is under neuronal regulation by SP neurons.


Similarities between crystal protein subunits of Bacillus thuringiensis strain 1884 serotype H14 and strain PG14 serotype H8a,8b, and their relationship with mosquitocidal activity.
Bacillus thuringiensis subsp. israelensis strain 1884 and B. thuringiensis subsp. morrisoni strain PG14 have the same toxicity toward mosquito larvae. Their protein crystal contents were compared either on native electrophoretic systems or on gel filtration. Analysis of electroeluted native proteins in SDS-PAGE showed that the 28-Kd protein alone, without the 68- or 130-Kd protein, was not larvicidal; however, toxicity was recorded as soon as 28- and 68-Kd proteins were associated. After gel filtration of 1884 soluble crystal, the 68-Kd protein alone, without 28 or 130 Kd, was not toxic; toxicity was also recorded when 28- and 68-Kd proteins and higher molecular weight proteins were associated. A comparable pattern was observed with PG14 soluble crystals.


Quantitative compensation by lateral motor column neurons in response to four functional hindlimbs in a frog tadpole.
An analysis of the lateral motor columns of a frog tadpole with 4 functional unilateral hindlimbs revealed a 58% increase in the number of neurons on the overloaded side as compared to the control side of the spinal cord. An accompanying, apparently selective, increase in the number of very large neurons suggests that spinal motor centers respond to peripheral overload through a variable combination of neuronal number and size in providing for adequate innervation.


Balloon-Expandable Pulmonary Valves for Patched or Native Right Ventricular Outflow Tracts.
The implantation of percutaneous balloon expandable valves in native or patched right ventricular outflow tracts (nRVOT) is a challenging technique due to the diversity of anatomies and shapes, the large sizes, and the distensibility of the nRVOT, for which specific techniques have been developed. We present a single center experience with balloon expandable percutaneous pulmonary valves in nRVOT, describing the techniques used, complications observed, and a short-mid term follow-up.. This is a single center descriptive study of patients who underwent a percutaneous pulmonary valve implantation in a nRVOT with a balloon expandable pulmonary valve in our center between September 2012 and June 2022.. We implanted successfully 45 valves in 46 patients (20 Sapien and 25 Melody). Tetralogy of Fallot or pulmonary atresia with VSD were the main congenital heart disease (n = 32). All were pre-stented, 18 in a one step procedure. We used a Dryseal sheath in 13/21 Sapien. In 6 patients we used the anchoring technique, 5 with a very large nRVOT and one pyramidal nRVOT. In the 3.5 year follow-up 7 patients developed endocarditis and 3 required a valve redilation, no fractures were observed. PPVI of native RVOT with balloon expandable valves is feasible in a number of selected anatomies, including large or pyramidal nRVOT, using specific techniques, (presenting, LPA anchoring).


Regulation of cytochrome P-450 CYPIA1 gene expression and proto-oncogene expression by growth factors in primary hepatocytes.
The effect of growth factors on the cytochrome P-450 (CYPIA1) gene expression was studied in primary mouse hepatocytes. Of the three growth factors used, i.e. epidermal growth factor (EGF), transforming growth factor alpha (TGF alpha) and insulin, only EGF or TGF alpha completely blocked CYPIA1 expression in the presence of the CYPIA1 inducer 3-methylcholanthrene (3-MC). This repression was not linked to cell cycle progression of the hepatocyte because insulin was active to induce 'early immediate genes' and DNA replication as well as EGF/TGF alpha but failed to suppress CYPIA1 expression. A specific EGF/TGF alpha receptor-mediated function may repress CYPIA1 gene expression and contribute to the acquisition of a xenobiotic drug resistance phenotype.


Developmental toxicity of Nafion byproduct 2 (NBP2) in the Sprague-Dawley rat with comparisons to hexafluoropropylene oxide-dimer acid (HFPO-DA or GenX) and perfluorooctane sulfonate (PFOS).
Nafion byproduct 2 (NBP2) is a polyfluoroalkyl ether sulfonic acid that was recently detected in surface water, drinking water, and human serum samples from monitoring studies in North Carolina, USA. We orally exposed pregnant Sprague-Dawley rats to NBP2 from gestation day (GD) 14-18 (0.1-30 mg/kg/d), GD17-21, and GD8 to postnatal day (PND) 2 (0.3-30 mg/kg/d) to characterize maternal, fetal, and postnatal effects. GD14-18 exposures were also conducted with perfluorooctane sulfonate (PFOS) for comparison to NBP2, as well as data previously published for hexafluoropropylene oxide-dimer acid (HFPO-DA or GenX). NBP2 produced stillbirth (30 mg/kg), reduced pup survival shortly after birth (10 mg/kg), and reduced pup body weight (10 mg/kg). Histopathological evaluation identified reduced glycogen stores in newborn pup livers and hepatocyte hypertrophy in maternal livers at ≥ 10 mg/kg. Exposure to NBP2 from GD14-18 reduced maternal serum total T3 and cholesterol concentrations (30 mg/kg). Maternal, fetal, and neonatal liver gene expression was investigated using RT-qPCR pathway arrays, while maternal and fetal livers were also analyzed using TempO-Seq transcriptomic profiling. Overall, there was limited alteration of genes in maternal or F1 livers from NBP2 exposure with significant changes mostly occurring in the top dose group (30 mg/kg) associated with lipid and carbohydrate metabolism. Metabolomic profiling indicated elevated maternal bile acids for NBP2, but not HFPO-DA or PFOS, while all three reduced 3-indolepropionic acid. Maternal and fetal serum and liver NBP2 concentrations were similar to PFOS, but ∼10-30-fold greater than HFPO-DA concentrations at a given maternal oral dose. NBP2 is a developmental toxicant in the rat, producing neonatal mortality, reduced pup body weight, reduced pup liver glycogen, reduced maternal thyroid hormones, and altered maternal and offspring lipid and carbohydrate metabolism similar to other studied PFAS, with oral toxicity for pup loss that is slightly less potent than PFOS but more potent than HFPO-DA.


Theoretical model for the post-transcriptional regulation of the human c-myc gene expression, involving double-stranded RNA processing.
We propose that the human c-myc gene expression is regulated by a post-transcriptional mechanism based on the formation of a hairpin structure between c-myc mRNA exons 1 and 2. This structure could be rapidly processed by a hypothetical double stranded RNAse, removing the 5' end segment of the c-myc mRNA having the cap site. Removal of cap should propitiate exonucleolytic degradation of transcript.


Distinct Proteins in Protein Corona of Nanoparticles Represent a Promising Venue for Endogenous Targeting - Part I: In vitro Release and Intracellular Uptake Perspective.
Protein corona (PC) deposition on nanoparticles (NPs) in biological systems contributes to a great extent to NPs' fates; their targeting potential, the interaction with different biological systems and the subsequent functions. PC - when properly tuned - can serve as a potential avenue for optimization of NPs' use in cancer therapy. Poly-lactic co-glycolic acid (PLGA)-based NPs exhibiting different physicochemical properties were fabricated and characterized. The PC makeup of these NPs were qualitatively and quantitatively analyzed by Western blot and Bradford assay, respectively. The effect of PC on the release of NPs' cargos and the intracellular uptake into B16F10 melanoma cells has been studied. The composition of NPs (polymeric PLGA NPs vs lipid-polymer hybrid NPs) and the conjugation of an active targeting ligand (cRGDyk peptide) represented the major determinants of the PC makeup of NPs. The in vitro release of the loaded cargos from the NPs depended on the PC and the presence of serum proteins in the release medium. Higher cumulative release has been recorded in the presence of proteins in the case of peptide conjugated NPs, cNPs, while the unconjugated formulations, uNPs, showed an opposite pattern. NPs intracellular uptake studies revealed important roles of distinct serum and cellular proteins on the extent of NPs' accumulation in melanoma cells. For example, the abundance of vitronectin (VN) protein from serum has been positively related to the intracellular accumulation of the NPs. Careful engineering of nanocarriers can modulate the recruitment of some proteins suggesting a potential use for achieving endogenous targeting to overcome the current limitations of targeted delivery of chemotherapeutic agents.


A randomized, controlled trial of a clinical pharmacist intervention in microdiscectomy surgery - Low dose intravenous ketamine as an adjunct to standard therapy.
The hypothesis that postoperative pain would be reduced by using 1 μg/kg/min of ketamine, both intra- and post-operatively, for lumbar microdiscectomy surgery was assessed by measuring morphine consumption. Patient side effects were reported. Forty-five patients undergoing microdiscectomy surgery were randomized under double-blind conditions into three groups: Group1 (G1) received normal saline, Group 2 (G2) ketamine (1 μg/kg/min) intra-operatively and Group 3 (G3) ketamine (1 μg/kg/min) both intra- and post-operatively. Morphine consumption, pain scores, nausea and vomiting, CNS disorders were recorded for 24 h post surgery. This study was conducted by applying the concept of a clinical pharmacist intervention. The time for the first analgesia demand dose was significantly shorter (P < 0.05) in G117 ± 1.7 min than for G2 and G3. In G3 morphine consumption 6, 12, and 24 h after surgery was 3 ± 2.26, 9.2 ± 2.11 and 26.9 ± 2.71 mg. Total morphine consumption was significantly lower for G3 than for G1 or G2 (P < 0.05). The visual analog scale score (VAS) values were significantly lower in G3 (P < 0.05) than for the other groups during the first 24 h. The rate of nausea and vomiting was significantly higher in G1 vs G3 (P < 0.05). No difference in drug induced CNS disturbances was observed among the groups. Using 1 μg/kg/min of ketamine hydrochloride intra- and post-operatively for microdiscectomy surgery could be an adjunct therapy to reduce postoperative morphine consumption minimizing its side effects. Collaborative clinical pharmacy practice on the basis of pharmacology had an effective role in improving the general outcome of microdiscectomy surgery.


Planned home birth assistance: challenges during the COVID-19 pandemic.
To understand the challenges faced by urban midwives in assisting planned home births during the COVID-19 pandemic. Qualitative study, based on the Collective Subject Discourse methodological framework, carried out with eight professionals, members of a birth care collective from the northeast region of Brazil. Data was collected between September and October of 2020 using the focus group technique. The collective discourses revealed five central ideas: Changing assistance strategy; Dealing with frustration; Facing the fear of contamination; Avoiding exposure to the virus; and Keeping distance during the care process. The challenging condition the pandemic brings to the care of planned home births is made evident, being marked by the need for collective protection and the pressure of following health recommendations. The study also points out the need for official protocols and good quality information based on scientific evidence and humanizing principles to guide health care.


Effects of SGM Education for Undergraduate Medical Students in a Canadian Context.
Medical schools have been striving to equip students with the tools and skills needed to serve patients from the LGBTQ + community, also called the Sexual and Gender Minority (SGM) community. This study aims to assess student comfort with providing care, and faculty knowledge and preparedness in delivering SGM-centered education at our home institution. We conducted two mixed-methods surveys, one geared towards medical students across four years of study and one towards medicine faculty. Each survey collected first demographic information about participants, then used a validated tool to assess knowledge of the SGM community. The qualitative component of both surveys then consisted of a needs assessment to determine what students felt should be changed about their curriculum, and what faculty felt should change about their training to deliver this curriculum. We received 26 student responses from all 4 years of study and 35 faculty responses from a variety of medical specialties. Difference in knowledge assessment scores was not statistically significant across both cohorts. Most students felt overall comfortable providing care for sexual minority individuals, and faculty similarly felt comfortable teaching, but data showcases that perceived comfort is higher among the student cohort. We propose that students are acquiring knowledge and comfort with providing for SGM individuals from sources outside their curriculum, and that additional training of faculty is vital to ensure students not doing this independent learning do not fall through the cracks. The online version contains supplementary material available at 10.1007/s40670-023-01831-x.


Process Development of Recombinant Adeno-Associated Virus Production Platform Results in High Production Yield and Purity.
Optimization of recombinant adeno-associated virus (rAAV) production has important clinical implications, as manufacturing is one of the major challenges for rAAV gene therapy. In this study, we optimized upstream and downstream processing of the rAAV production platform created by an earlier design-of-experiment approach. Our results showed that adding peptones (yeastolate, Trypton N1 or both) increased production yield by 2.8- to 3.4-folds. For downstream processing, a variety of wash buffers for an affinity resin, POROS™ CaptureSelect™ (PCS)-AAVX, were tested for their effects on rAAV8 purity, including NaCl, MgCl2, arginine, Triton X-100, CHAPS, Tween 20, octyl β-d-1-thioglucopyranoside (OTG), and low pH. The results showed that the OTG wash significantly improved the rAAV purity to 97% and reduced endotoxins to an undetectable level (<0.5 EU/mL), while retaining the yield at 92.3% of the phosphate-buffered saline (PBS) wash. The OTG wash was successfully applied to purifications of rAAV1, rAAV2, and rAAV5 using PCS-AAVX, and rAAV9 using PCS-AAV9. rAAV8 purified with OTG wash showed comparable transduction efficiency in HEK 293T cells to the rAAV8 purified with PBS wash. The optimized rAAV production process yielded 5.5-6.0 × 1014 and 7.6 × 1014 vector genome per liter of HEK 293T cells for purified rAAV8- and rAAV5-EF1α-EGFP (enhanced green fluorescent protein), respectively. The platform described in this study is simple with high yields and purity, which will be beneficial to both research and clinical gene therapy.


Sustained morphine treatment augments basal CGRP release from cultured primary sensory neurons in a Raf-1 dependent manner.
Recent studies suggest that sustained morphine-mediated paradoxical pain may play an important role in the development of analgesic tolerance. The intracellular signal transduction pathways involved in sustained opioid mediated augmentation of spinal pain neurotransmitter (such as calcitonin gene-related peptide (CGRP)) release are not fully clarified. Cyclic AMP (cAMP)-dependent protein kinase (PKA) plays an important role in the modulation of presynaptic neurotransmitter release. Moreover, we have shown earlier that sustained opioid agonist treatment leads to a Raf-1-dependent sensitization of adenylyl cyclase(s) (AC superactivation), augmenting forskolin-stimulated cAMP formation upon opioid withdrawal (cAMP overshoot). Therefore, in the present study we examined the role of Raf-1 in sustained morphine-mediated regulation of cAMP formation and basal CGRP release in vitro, in cultured neonatal rat dorsal root ganglion (DRG) neurons. We found that sustained morphine treatment significantly augments intracellular cAMP production as well as basal CGRP release from cultured neonatal rat DRG neurons. The selective PKA inhibitor, H-89, attenuates the sustained morphine-mediated augmentation of basal CGRP release, indicating that the cAMP/PKA pathway plays an important role in regulation of CGRP release from sensory neurons. Since our present data also demonstrated that selective Raf-1 inhibitor, GW 5074, attenuated both the cAMP overshoot and the augmentation of CGRP release mediated by sustained morphine in neonatal rat DRG neurons, we suggest that Raf-1-mediated sensitization of the intracellular cAMP formation may play an important role in sustained morphine-mediated augmentation of spinal pain neurotransmitter release.


Sensory and motor responses of trigeminal and reticular neurons during ingestive behavior in rats.
Activities of 53 neurons in the brain stem were recorded with chronically implanted fine wires in freely eating and drinking rats. Twenty units were isolated from the trigeminal mesencephalic nucleus; 18 were spindle afferents and 2 periodontal afferents. The spindle units were classified into 4 types: 5 units showed rhythmical activity related only to the jaw opening phase during both licking and chewing, 8 units discharged at jaw opening phase during licking, but both at jaw opening and jaw closing phases during eating, 2 units increased phasic activity at jaw opening phase during licking, but increased tonically independent of jaw movements during eating, and the remaining 3 units responded only at jaw closing phase both in licking and eating behavior. Nine units were assumed to be alpha motoneurons isolated from the trigeminal motor nucleus; 2 innervating the temporalis muscle, 3 the masseter muscle, 2 the digastric muscle, and the remaining 2 presumably the pterygoid muscles. These units, with a mean tonic spontaneous rate of about 10 impulses/s, showed phase-related rhythmical burst activities during licking and eating. Four units were isolated from the trigeminal main sensory nucleus. Sixteen units were isolated from the medullary reticular formation and 1 from the pontine reticular formation. Of these 16 units, 3 from the parvocellular part of the medullary reticular formation were presumably premotor interneurons, and 3 units in the "intertrigeminal region", 2 units in the "juxtatrigeminal region", and 8 units in the "supratrigeminal region" were responsive to multiple sensory modalities. These results demonstrate that almost all the units sampled showed characteristic rhythmic activities associated with the position of the tongue or mandible during rhythmical jaw movements.


Potassium-39 nuclear magnetic resonance observation of intracellular potassium without chemical shift reagents during metabolic inhibition in the isolated perfused rat heart.
The intracellular potassium content of perfused rat heart was measured by potassium-39 nuclear magnetic resonance (NMR) spectroscopy at 33 degrees C with an inversion recovery technique based on the fact that the spin-lattice relaxation time (T1) of the intracellular potassium (8.3 msec at 8.45 T) is much faster than that of the extracellular potassium (68 msec). Intracellular potassium decreased to 60.2 +/- 4.3% of the control level (mean +/- SEM, n = 6) at 40 minutes from the start of metabolic inhibition (2 mM cyanide, 0 mM glucose). Removal of cyanide restored intracellular potassium to 94.2 +/- 3.9% at 30 minutes from the restart of oxidative metabolism. The cumulative potassium loss was determined from the flow rate and potassium concentration of the coronary effluent, which reached 139 +/- 12 mumol/g dry wt during 40 minutes of metabolic inhibition. This value was calculated as 41.8% of intracellular potassium in the control heart and agreed with the decrement of intracellular potassium measured by NMR. During the metabolic inhibition and recovery period, a linear correlation was observed between the changes in 39K NMR-observed intracellular potassium and the cumulative potassium loss. The present results evaluate the inversion recovery technique as a method to successfully monitor the myocardial intracellular potassium.


Amino acid residues important for D-galactose recognition by the F-type lectin, Ranaspumin-4.
F-type lectins are typically L-fucose binding proteins with characteristic L-fucose-binding and calcium-binding sequence motifs, and an F-type lectin fold. An exception is Ranaspumin-4, an F-type lectin of the Tungra frog, Engystomops pustulosus. Ranaspumin-4 is D-galactose specific and does not bind to L-fucose although it has the conserved L-fucose binding sequence motif and shares overall sequence similarity with other F-type lectins. Here, we report the detailed glycan-binding profile of wild-type Ranaspumin-4 using hemagglutination inhibition assays, flow cytometry assays and enzyme-linked lectin assays, and identify residues important for D-galactose recognition using rational site-directed mutagenesis. We demonstrate that Ranaspumin-4 binds to terminal D-galactose in α or β linkage with preference for α1-3, α1-4, β1-3, and β1-4 linkages. Further, we find that a methionine residue (M31) in Ranaspumin-4 that occurs in place of a conserved Gln residue (in other F-type lectins), supports D-galactose recognition. Resides Q42 and F156 also likely aid in D-galactose recognition.


Relationship between vitamin D and asthma from gestational to adulthood period: a meta-analysis of randomized clinical trials.
Despite numerous studies investigating vitamin D, its impact on asthma is still unknown. The aim of our meta-analysis is to analyze the vitamin D supplementation influence on asthma prevention and treatment ranging from gestational to adulthood period. Fifteen randomized clinical trials were included after database search. Studies contained the analyzed endpoints: the number of asthma and wheezing occurrence in gestational and infantile periods, the change of childhood/adult asthma control test score and forced expiratory volume in one second (FEV1) in childhood and adulthood periods. Random effects model was used to calculate effect sizes. Supplementation by women during pregnancy period decreased the wheezing occurrence in their children by 23% (RR = 0.77; 95% CI [0.64; 0.92]; p < 0.0049, I2 = 0%); whereas had no effect on given asthma parameters during the infantile period. Moreover, vitamin D administration had negative effect on the FEV1 change in children (MD = -3.84; 95% CI [-7.68; -0.01]; p = 0.0497; I2 = 95%), but had positive effect on the change of ACT score in adults (MD = 1.80; 95% CI [0.12; 3.49]; p = 0.0359; I2 = 99%). Our meta-analysis showed the varying results depending on patient's life period. It is important to further investigate the role of vitamin D supplementation in asthma management.


Expression level of PD-L1 is involved in ALDH1A1-mediated poor prognosis in patients with head and neck squamous cell carcinoma.
To evaluate the expression levels of ALDH1A1, PDL1, and PDL2 in head and neck squamous cell carcinoma (HNSCC) patients, and explore their clinical relevance in prognosis of patients with HNSCC. Immunohistochemistry of ALDH1A1 and PD-L1/PD-L2 in 85 primary HNSCC patients was carried out. The expression level of PD-L2 was assessed with the modified Moratin's immune response scoring (IRS) system. tumor proportion score (TPS) was defined as the percentage of viable tumor cells showing partial or complete membrane staining at any intensity. The chi-square test and Fisher's exact test were used to analyze the associations between ALDH1A1 expression and clinicopathological features. The Spearman's correlation was applied to analyze the correlation of ALDH1A1 expression with PD-L1/PD-L2 expression. kaplan-Meier analysis showed that the expression levels of ALDH1A1 and PD-L1/PD-L2 were inversely associated with recurrence-free survival (RFS; P = 0.001, 0.014, and 0.023, respectively). Moreover, expression levels of ALDH1A1 and PD-L1 were correlated with poor overall survival (OS; P = 0.002 and 0.039, respectively). Furthermore, multivariate logistics regression analyses demonstrated that expression level of ALDH1A1 was independently associated with shorter RFS (P = 0.013) and poorer OS (P = 0.014) in HNSCC patients, and the expression level of PD-L2 was only negatively associated with RFS (P = 0.041), rather than PD-L1. Spearman's correlation analysis unveiled that expression levels of PD-L1 and PD-L2 were positively correlated with ALDH1A1 expression in HNSCC patients (P = 0.000 and 0.015, respectively). Especially, the patients with expression levels of ALDH1A1 and PD-L1 had the worst prognosis. Our results indicated that ALDH1A1 is an independent prognostic factor in patients with HNSCC, and the expression level of PDL-1 may be involved in ALDH1A1-mediated poor prognosis in patients with HNSCC.


Changes in self-efficacy and dietary adherence: the impact on weight loss in the PREFER study.
Findings from studies examining self-efficacy and its relationship to weight loss have been inconsistent. We examined self-efficacy specific to changing eating behaviors in the PREFER trial, an 18-month behavioral weight-loss study, to determine if self-efficacy and dietary adherence were associated with weight change, and what impact self-efficacy had on weight change after controlling for adherence. Measurements included the weight efficacy lifestyle (WEL) questionnaire, body weight, self-reported fat gram intake, kilocalorie intake, and adherence to kilocalorie and fat gram goals at baseline, 6, 12, and 18 months. The sample (N = 170) was 88.2% female and 70.0% Caucasian; the mean age was 44.1 years (SD = 8.8). Mean weight loss at 18 months was 4.64% (SD = 6.24) of baseline body weight and the mean increase in self-efficacy was 11.70% (SD = 38.61). Self-efficacy improved significantly over time (p = 0.04) and was associated with weight loss (p = 0.02). Adherence to the fat gram goal was associated with weight loss (p = 0.0003), and self-efficacy remained associated with weight loss after controlling for fat gram adherence (p = 0.0001). Consistent with self-efficacy theory, improvement in self-efficacy over time supported greater weight loss. Adherence to the fat gram goal also influenced weight loss.


Humoral serological response to the BNT162b2 vaccine after allogeneic haematopoietic cell transplantation.
To assess the humoral immune response to the BNT162b2 vaccine after allogeneic haematopoietic cell transplantation (HCT). This is a prospective cohort study. The SARS-CoV-2 IgGII Quant (Abbott©) assay was performed 4-6 weeks after the second BNT162b2 vaccine for quantitative measurement of anti-spike antibodies. The cohort included 106 adult patients. Median time from HCT to vaccination was 42 (range 4-439) months. Overall, 15/106 (14%, 95% confidence interval (CI) 7-21%) were seronegative despite vaccination, 14/52 patients on immunosuppression (27%, 95%CI 19-35%) compared to only 1/54 patients off immunosuppression (1.8%, 95%CI 1-4%) (p 0.0002). The proportion of seronegative patients declined with time; it was 46% (6/13) during the first year, 12.5% (3/24) during the second year and 9% (6/69) beyond 2 years from transplant. Patients with acute graft-versus-host disease (GVHD) (odds ratio (OR) 3.3, 95%CI 0.97-11.1, p 0.06) and moderate to severe chronic GVHD (OR 5.9, 95%CI 1.2-29, p 0.03) were more likely to remain seronegative. Vaccination was well tolerated by most patients. However, 7% (7/106) reported that GVHD-related symptoms worsened within days following vaccination. A significant proportion of allogeneic HCT recipients receiving immunosuppression demonstrated an inadequate humoral response to the BNT162b2 vaccine. These patients should be recognized and instructed to take appropriate precautions. Recipients who were off immunosuppression had a humoral response that was comparable to that of the general population.


Development of a simple assay system for protein-stabilizing efficiency based on hemoglobin protection against denaturation and measurement of the cooperative effect of mixing protein stabilizers.
We have elucidated the cooperative stabilization of proteins by sugars, amino acids, and other protein-stabilizing agents using a new and simple assay system. Our system determines the protein-stabilizing ability of various compounds by measuring their ability to protect hemoglobin from denaturation. Hemoglobin denaturation was readily measured by quantitative changes in its ultraviolet-visible absorption spectrum. The efficiency of our assay was confirmed using various sugars such as trehalose and sucrose that are known to be good protein stabilizers. We have also found that mixtures of two different types of protein stabilizers resulted in a cooperative stabilizing effect on protein.


Financial Cost of Hypertension in Urban and Rural Tertiary Health Facilities in Southwest, Nigeria: A Comparative Cross-Sectional Study.
The financial cost of hypertension could result in serious economic hardship for patients, their households, and the community. To assess and compare the direct and indirect cost of care for hypertension in urban and rural tertiary health facilities. A comparative cross-sectional study was carried out in two tertiary health facilities which are located in urban and rural communities of the southwest, Nigeria. Four hundred and six (204 urban, 202 rural) hypertensive patients were selected from the health facilities using a systematic sampling technique. A pretested semi-structured, interviewer-administered questionnaire adapted from that used in a previous study was used for data collection. Information on biodata, and direct and indirect costs was collected. Data entry and analysis were done using IBM SPSS Statistics for Windows, Version 22.0. More than half of the respondents were females (urban, 54.4%; rural, 53.5%) and in their middle age (45-64 years) (urban, 50.5%; rural, 51.0%). The monthly cost of care for hypertension was significantly higher in urban than in rural tertiary health facilities (urban, 19,703.26 [$54.73]; rural, 18,448.58 [$51.25]) (P < 0.001). There was a significant difference in the direct cost (urban, 15,835.54 [$43.99]; rural, 14,531.68 [$40.37]) (P < 0.001), although the indirect cost (urban, 3,867.72 [$10.74]; rural, 3,916.91 [$10.88]) (P = 0.540) did not show much difference between the groups. The cost of drugs/consumables and investigations contributed more than half (urban, 56.8%; rural, 58.8%) of the cost in both health facilities. The financial cost of hypertension was higher in the urban tertiary health facility; therefore, more government support is needed in this health facility to close the financial gap.


Spontaneous attachment of detached donor-corneal graft following Descemet's stripping automated endothelial keratoplasty.
This case report presents a case that had Descemet's stripping automated endothelial keratoplasty complicated postoperatively by detached donor grafts. The patient had a well centered partially detached graft that attached spontaneously the next day without the need to re-bubble. Postoperatively he had a clear cornea, improved uncorrected visual acuity with good endothelial cell count. It is worth waiting for a partially detached well centered graft to adhere spontaneously; this would lower the possible risks of repeated anterior chamber air injections. Cases should be selected to eliminate any mechanical factor that would prevent the attachment.


Recurrence Rate and Relevant Associated Factors of Stroke among Patients with Small Artery Occlusion in Northern China.
Small artery occlusion (SAO) is responsible for 31.3% of all ischemic strokes in China. However, reports regarding the recurrence rate of SAO in China are rare. We aimed to assess the recurrence rate and factors associated with SAO in China. All consecutive patients with SAO hospitalized at Tianjin Huanhu Hospital from 2005 to 2014 were recruited. We assessed the association between stroke subtype, severity, and disease history with recurrence at 3, 12, and 36 months of onset using multivariate logistic regression analysis. A total of 2,524 SAO patients were included in this study, including 1696 (67.2%) men and 828 (32.8%) women. The recurrence rates were 3.1% at 3 months, 12.7% at 12 months, and 36.5% at 36 months. Compared with women, men had a higher risk of recurrence at 3 months after SAO (P = 0.003). Old age and severity of stroke were also associated with a higher risk of recurrence (P < 0.05). Patients with an elevated C-reactive protein had a higher risk of recurrence at 12 months (P = 0.003). On the other hand, the risk of recurrence at 12 months was 39% lower in patients who consumed alcohol than in those who did not (P = 0.037). Hypertension, atrial fibrillation, and obesity were independent risk factors of recurrence at 36 months. These findings suggest that modification of risk factors in patients with SAO, particularly men, is essential for reducing the rate of recurrence and the overall burden of stroke in China.


Longan pulp polysaccharides regulate gut microbiota and metabolites to protect intestinal epithelial barrier.
Longan pulp polysaccharide is a bioactive component with prebiotic activity and intestinal barrier protection. This study aimed to evaluate the influence of digestion and fermentation on the bioavailability and intestinal barrier protection of polysaccharide LPIIa from longan pulp. The molecular weight of LPIIa didn't change significantly after gastrointestinal digestion in vitro. After fecal fermentation, 56.02% of LPIIa was consumed by gut microbiota. The short-chain fatty acid level in LPIIa group was 51.63% higher than that in blank group. LPIIa intake also increased short-chain fatty acid production and G-protein-coupled receptor 41 expression in the colon of mice. Moreover, LPIIa improved the relative richness of Lactobacillus, Pediococcus, and Bifidobacterium in colon content. Compared to LPIIa, fecal fermented LPIIa better protected intestinal epithelial barrier by increasing Zonula occludens-1 expression. These results provided an important basis for the design of functional food based on longan polysaccharides to prevent intestinal barrier damage related diseases.


Metabolic environment in vivo as a blueprint for differentiation and maturation of human stem cell-derived cardiomyocytes.
Patient-derived human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are increasingly being used for disease modeling, drug screening and regenerative medicine. However, to date, an immature, fetal-like, phenotype of hPSC-CMs restrains their full potential. Increasing evidence suggests that the metabolic state, particularly important for provision of sufficient energy in highly active contractile CMs and anabolic and regulatory processes, plays an important role in CM maturation, which affects crucial functional aspects of CMs, such as contractility and electrophysiology. During embryonic development the heart is subjected to metabolite concentrations that differ substantially from that of hPSC-derived cardiac cell cultures. A deeper understanding of the environmental and metabolic cues during embryonic heart development and how these change postnatally, will provide a framework for optimizing cell culture conditions and maturation of hPSC-CMs. Maturation of hPSC-CMs will improve the predictability of disease modeling, drug screening and drug safety assessment and broadens their applicability for personalized and regenerative medicine.


Relating Retinal Morphology and Function in Aging and Early to Intermediate Age-related Macular Degeneration Subjects.
To evaluate relationships between age-related macular degeneration (AMD) morphology on spectral-domain optical coherence tomography (SDOCT) and visual function. Cross-sectional, observational. From the Alabama Study on Early AMD baseline visit, visual acuity, cone-mediated sensitivity, rod-mediated dark adaptation, and SDOCT were obtained in 1 eye per subject with no apparent retinal aging (n = 15), normal aging (n = 15), early AMD (n = 15), and intermediate AMD (n = 46). The volumes of retinal pigment epithelium (RPE)-drusen complex, RPE-drusen complex abnormal thinning, RPE-drusen complex abnormal thickening, and inner and outer retina were calculated in specified regions using semi-automated SDOCT segmentation. Better cone-mediated sensitivity was associated with greater RPE-drusen complex volume (r = 0.34, P < .001) and less RPE-drusen complex abnormal thinning volume (r = -0.31, P = .003). Longer rod-mediated dark adaptation time, the duration for rod-mediated sensitivity to recover from photo-bleach exposure, correlated with lower RPE-drusen complex volume (r = -0.34, P = .005) and greater RPE-drusen complex abnormal thinning volume (r = 0.280, P = .023). In 19 eyes with subretinal drusenoid deposits (SDD) vs 47 eyes without SDD, rod-mediated dark adaptation time was longer (mean ± SD 13.5 ± 7.0 vs 10.2 ± 3.1 minutes, P = .004), RPE-drusen complex abnormal thinning volume was greater (P < .0001), and visual acuity and cone sensitivity did not differ. Decreased function relates to structural markers on SDOCT in AMD. Because the RPE-drusen complex includes the interdigitation of outer segments and RPE apical processes and SDD in eyes with AMD, slower dark adaptation might be related to structural abnormalities of the RPE, the RPE-photoreceptor interface, or both.


Lower Total Knee Arthroplasty Revision Risk Associated With Bisphosphonate Use, Even in Patients With Normal Bone Density.
Bisphosphonates (BPs) are associated with lower total knee arthroplasty (TKA) revision risk, but the effect of bone mineral density has not been evaluated. A cohort of 34,116 primary TKA patients was evaluated with revision surgery and periprosthetic fractures as end points. BP usage was the exposure of interest. Bone quality (normal, osteopenia, and osteoporosis) and patient age (<65 vs ≥65 years) were evaluated as effect modifiers of risk estimates. Of the patients, 19.6% were BP users. In BP users, 0.5% underwent an aseptic revision; and 0.6%, a periprosthetic fracture. In non-BP users, 1.6% underwent aseptic revision; and 0.1%, a periprosthetic fracture. Bisphosphonate use was associated with lower risk of revision in all bone quality categories in those older than 65 years. The risk of periprosthetic fractures was higher for patients on BP.


GPR64, Screened from Ewing Sarcoma Cells, Is a Potential Target for Antibody-Based Therapy for Various Sarcomas.
Ewing sarcoma is an aggressive and the second most common bone tumor in adolescent and young adult patients. The 5-year survival rate is 60-70% for localized disease but 30% for patients with metastases. Here, we aimed to identify a therapeutic target for Ewing sarcoma and evaluate antibody-based therapeutic agents using in vitro and in vivo models. We identified G protein-coupled receptor 64 (GPR64) as a therapeutic target for Ewing sarcoma via next-generation RNA-sequencing. GPR64v205 mRNA was expressed in HTB166, A673, MG63, 143B, HS-Sy II, and HT1080 cell lines as well as in Ewing sarcoma, undifferentiated pleomorphic sarcoma, leiomyosarcoma, dedifferentiated liposarcoma, and synovial sarcoma tissues. GPR64 expression was observed in 62.5% of sarcoma cases and was overexpressed in 33.9% cases. GPR64-specific monoclonal antibodies were tested as near-infrared probes for in vivo imaging using subcutaneous tumor mouse xenografts. Fluorescence intensity was stronger for the AF700-labeled anti-GPR64 antibody than that for the AF700-labeled isotype control antibody. GPR64 was detected in engrafted tumors of A673, 143B, HT1080, and the epididymis but not in other resected tissues. The anti-GPR64 antibody showed excellent binding to GPR64-positive tumors but not to healthy tissues. This antibody has potential for drug delivery in the antibody-based treatment of sarcomas.


Influent wastewater microbiota and temperature influence anaerobic membrane bioreactor microbial community.
Sustainable municipal wastewater recovery scenarios highlight benefits of anaerobic membrane bioreactors (AnMBRs). However, influences of continuous seeding by influent wastewater and temperature on attached-growth AnMBRs are not well understood. In this study, four bench-scale AnMBR operated at 10 and 25°C were fed synthetic (SPE) and then real (PE) primary effluent municipal wastewater. Illumina sequencing revealed different bacterial communities in each AnMBR in response to temperature and bioreactor configuration, whereas differences were not observed in archaeal communities. Activity assays revealed hydrogenotrophic methanogenesis was the dominant methanogenic pathway at 10°C. The significant relative abundance of Methanosaeta at 10°C concomitant with low acetoclastic methanogenic activity may indicate possible Methanosaeta-Geobacter direct interspecies electron transfer. When AnMBR feed was changed to PE, continual seeding with wastewater microbiota caused AnMBR microbial communities to shift, becoming more similar to PE microbiota. Therefore, influent wastewater microbiota, temperature and reactor configuration influenced the AnMBR microbial community.


Multi-Layered TiO₂ Films towards Enhancement of Escherichia coli Inactivation.
Crystalline TiO₂ has shown its great photocatalytic properties in bacterial inactivation. This work presents a design fabrication of low-cost, layered TiO₂ films assembled reactors and a study of their performance for a better understanding to elucidate the photocatalytic effect on inactivation of E. coli in water. The ability to reduce the number of bacteria in water samples for the layered TiO₂ composing reactors has been investigated as a function of time, while varying the parameters of light sources, initial concentration of bacteria, and ratios of TiO₂ film area and volume of water. Herein, the layered TiO₂ films have been fabricated on the glass plates by thermal spray coating prior to screen printing, allowing a good adhesion of the films. Surface topology and crystallographic phase of TiO₂ for the screen-printed active layer have been characterized, resulting in the ratio of anatase:rutile being 80:20. Under exposure to sunlight and a given condition employed in this study, the optimized film area:water volume of 1:2.62 has shown a significant ability to reduce the E. coli cells in water samples. The ratio of surface area of photocatalytic active base to volume of water medium is believed to play a predominant role facilitating the cells inactivation. The kinetic rate of inactivation and its behavior are also described in terms of adsorption of reaction species at different contact times.


High-mobility group box-1 peptide ameliorates bronchopulmonary dysplasia by suppressing inflammation and fibrosis in a mouse model.
This study aimed to examine the effect of the HMGB1 peptide on Bronchopulmonary dysplasia (BPD)-related lung injury in a mouse model. HMGB1 peptide ameliorates lung injury by suppressing the release of inflammatory cytokines and decreasing soluble collagen levels in the lungs. Single-cell RNA sequencing showed that the peptide suppressed the hyperoxia-induced inflammatory signature in macrophages and the fibrotic signature in fibroblasts. These changes in the transcriptome were confirmed using protein assays. Systemic administration of HMGB1 peptide exerts anti-inflammatory and anti-fibrotic effects in a mouse model of BPD. This study provides a foundation for the development of new and effective therapies for BPD.


Sorption and desorption characteristics of anionic surfactants to soil sediments.
Surfactants are important environmental chemicals due to their extensive domestic and industrial applications, such as subsurface organic pollution remediation and enhanced oil recovery. However, the interaction of surfactants with subsurface material particularly the desorption behavior of surfactants is less understood. Surfactant desorption is essential to control the fate and transport of surfactants as well as organic pollutants. In this study, the sorption and desorption of linear sodium dodecylbenzene sulfonate (SDBS) and sodium hexadecyl diphenyl oxide disulfonate (DPDS) with two types of soil sediment samples are compared. Sorption of surfactants can be modeled by hydrophobic sorption. Less DPDS sorption is observed at a higher aqueous concentration, which is attributed to the competition between surfactant micelles and sediment organic matter for DPDS sorption. A significant fraction of the sorbed surfactants resists desorption, and this is not a result of surfactant precipitation or desorption kinetics. Surfactant desorption behavior is similar to the irreversible desorption of hydrocarbons from soil with only half of the resistant phase surfactant being readily extracted by heated solvent extraction. The sorption/desorption data are interpreted with a molecular topology and irreversible sorption model. The knowledge of this study can be useful in understanding the environmental fate and transport of these common anionic surfactants. The methodology developed in this study can be expanded to study the sorptive nature of a wider range of surfactants in the environment.


Reactivity of ionic liquids with kaolinite: melt intersalation of ethyl pyridinium chloride in an urea-kaolinite pre-intercalate.
This paper reports a new route for the intercalation of an ionic liquid, namely 1-ethyl pyridinium chloride, into the interlamellar spaces of kaolinite. The intercalation was achieved using a kaolinite-urea intercalate as a starting material. The results of the XRD, FTIR, and TGA analyses confirmed the intercalation of ethyl pyridinium in the interlamellar spaces of kaolinite. 13C CP/MAS spectra indicated the complete displacement of urea by ethyl pyridinium. 29Si and 27Al NMR spectra of the starting materials and the products are also discussed as well as the results of the elemental analysis of the produced nanohybrid material.


Traumatic stress, mental health, and quality of life in adolescents with esophageal atresia.
We aimed to investigate QoL in EA patients in relation to comparison groups and to clinical factors including experienced traumatic stress. Adolescents with EA in Norway born between 1996 and 2002 were included. Clinical assessment and patient's characteristics were collected. Quality of life (PedsQL), traumatic stress (IES-13) and mental health (SDQ-20) were compared to groups of healthy controls, children with acute lymphoblastic leukemia (ALL) and kidney transplanted children (TX). 68 EA adolescents participated. Total scores for PedsQL were not different from the healthy group and ALL patients, but significantly better than the TX patients. The subscale for physical performance was significantly lower than in healthy adolescents, and nine (17%) patients had scores ≤70 indicating reduced health status. Five EA adolescents (12%) had mental health scores suggesting a psychiatric disorder, and six (9%) reported high traumatic stress scores with a significant correlation to days on ventilator in the neonatal period. The strongest predictors for quality of life among EA adolescents were self-reported mental health, posttraumatic stress and GERD symptoms. Scores for Quality of life in the EA group are good except for subscale for physical performance. Symptoms of posttraumatic stress, mental strain and gastroesophageal reflux are predictors of reduced QoL.


Current practice of care for adolescent and adult patients after Fontan surgery in Poland.
The growing number of adult patients after the Fontan operation requires regular surveillance tests in specialized centers. Our study aimed to evaluate the current practice of care for Fontan patients in Poland using a multicenter survey. Eight centers were included in the study including 5 adult congenital heart disease (ACHD) and 3 pediatric centers for adolescents. To compare the centers and facilitate interpretation of results, the Fontan Surveillance Score (FSS) was developed. The higher score is consistent with better care, with a maximum of 19 points. We included in the study 398 Fontan patients (243 adults and 155 adolescents [aged 14-18 years]). The median FSS was 13 points with variability between centers (interquartile range 7-14 points). Centers providing continuous care from the pediatric period until 18 years of age achieved a higher FSS compared to ACHD centers (median: 14 points vs. 12 points; P <0.001). Most of the patients, both in the ACHD (82.3%) and pediatric centers (89%), were seen annually and had a physical examination, electrocardiogram, and echocardiogram performed at each visit. However, we observed unsatisfactory utilization of tests identifying early stages of Fontan circulation failure (cardiopulmonary exercise tests, cardiac magnetic resonance, liver biochemistry and imaging, detection of protein-losing enteropathy). Our results showed that there is no unified surveillance approach for Fontan patients in Poland. The practice of care for adults differs from that of adolescents.


Peripheral T-cell lymphoma mimicking marginal zone B-cell lymphoma.
Peripheral T-cell lymphoma (PTCL) may assume a variety of histologic and cytologic appearances. We describe eight cases of PTCL morphologically simulating marginal zone B-cell lymphoma. We reviewed PTCL cases diagnosed in our institution between 1990 and 2000 and selected eight cases for study based on the following criteria: small-cell morphology with abundant, clear cytoplasm and either marginal zone involvement by the neoplastic infiltrate in lymph node biopsies or lymphoepithelial lesions in extranodal biopsies. Histologic features and ancillary studies were reviewed. Patients included six women and two men with a median age of 53 years (range, 35 to 74 years). Six patients were diagnosed with primary nodal PTCL, and two presented with primary extranodal disease. The original diagnosis was PTCL in only four cases; three cases were diagnosed as atypical lymphoid infiltrate, and one case as benign lymphoepithelial lesion. Lymph node biopsies revealed partial effacement of the architecture with residual follicles surrounded by the neoplastic small cells. Extranodal sites included hard palate, tongue, tonsil, and submandibular glands; all but one case demonstrated lymphoepithelial lesions. Monoclonality was demonstrated in six of eight cases (rearrangement of T-cell receptor gene), and three of eight had an aberrant T-cell population by flow cytometry. The differential diagnosis of atypical lymphoid infiltrates with morphologic features of marginal zone B-cell lymphoma should include PTCL. This uncommon morphological mimicry should be recognized, because PTCL is an aggressive disease regardless of morphology and should be treated accordingly.


Nanoencapsulation of anthocyanins-loaded β-lactoglobulin nanoparticles: Characterization, stability, and bioavailability in vitro.
This work aims to investigate the effect of desolvation on the stability and bioavailability of nanoparticles of β-lactoglobulin (β-Lg) and anthocyanins (AC) extracted from red raspberry pomace. Interactions between the substrates were also studied using multispectral approaches. β-Lg-nanoparticles were fabricated via heat treatment at 85 °C for 30 min before initiating the desolvation method at pH 7. This method generated monodisperse particles, nano-scale size of β-Lg, and AC-β-Lg ranged from 129.13 to 351.85 nm with square morphology obtained by SEM. The AC extract was encapsulated successfully during desolvation process into β-Lg-nanoparticles with encapsulation efficiency (EE %) of ~77%. Results also showed that AC (from 1 to 13 × 10-4 M) quenched the fluorescence intensity of de-solvated β-Lg estimated to be 98%, and a binding among them occurred with a Ka-value of 7.59 × 108 M-1 at 25 °C. Addition of AC also gradually increased the antioxidant activity of β-Lg-nanoparticles with values of 82.51% at the highest AC-concentration (13 × 10-4 M) loaded on β-Lg-nanoparticles. AC-loaded β-Lg nanoparticles was more stable in mouth (pH 6.8), simulated gastric (SG, pH 2), and simulated intestinal (SI, pH 6.9) by showing high retention rate (%) than that of AC unencapsulated. Overall, de-solvated-β-Lg increased the heat-stability and bioavailability of AC, which could be further utilized in various food and pharmaceutical matrices. These findings recommend that β-Lg nanoparticles could be appropriated as delivery systems for anthocyanins.


[Prescribing beta blockers in elderly patients with heart failure].
Beta blockers remain underused in elderly patients with heart failure. Age is not a contraindication to beta blockers. The SENIORS study confirmed that beta blockers are both efficacious and well tolerated in elderly people with heart failure, regardless of their ejection fraction. Because adverse effects may be both more frequent and more serious in the elderly, prescription protocols must be strictly applied. Patients in stable NYHA stages II or III may begin beta blocker treatment, at least 1 month after any decompensation. The initial dose must be as low as possible (1.25 mg/d for bisoprolol and nebivolol). Doses must be increased very progressively and stages longer than 15 days may be necessary. The objective is to reach the target dose (10mg/d for bisoprolol and nebivolol), given the dose-response effect that exists for beta blockers in elderly people with heart failure. In the case of low blood pressure, antihypertensive treatments must be reduced or stopped (for example, nitrate derivatives or calcium channel blockers). A reduction in the dosage of any diuretic dosage and finally of the beta blocker may follow, if necessary. Should bradycardia occur, any anti-bradycardia treatments (such as digoxin or amiodarone) must be reduced or stopped before the beta blocker dosage is reduced.


Interferon-gamma Immunotherapy in a Patient With Refractory Disseminated Candidiasis.
Despite advances in supportive care and novel antifungal agents, mortality caused by invasive Candida infection is high. A 3-year-old boy with disseminated Candida dubliniensis infection during induction chemotherapy for acute lymphoblastic leukemia deteriorated despite resolution of neutropenia and appropriate antifungal treatment. Monocyte human leukocyte antigen-DR expression was extremely low, suggesting immunoparalysis. Adjuvant immunotherapy with interferon-gamma restored the immune response, which was accompanied by clinical and radiographic recovery.


Inhibition of steroidogenic response to corticotropin in mouse adrenal tumor cells (Y-1) by the ionophore A23187. Role of protein biosynthesis.
Addition of the ionophore A23187 to Y-1 mouse adrenal tumor cells in monolayer culture inhibits steroidogenesis and the steroidogenic response to corticotropin (50% inhibition at 1 . 10(-7)M). Inhibition is rapid in onset and is not overcome by addition of external Ca2+. The ionophore also inhibits stimulation of steroid synthesis by cyclic AMP. A23187 inhibits incorporation of the amino acid lysine into protein by Y-1 cells and the dose dependence of this inhibition closely resembles that of the inhibition of the steroidogenic response to corticotropin. Addition of A23187 to a subcellular system for protein synthesis prepared from Y-1 cells, inhibits incorporation of the amino acid phenylalanine into protein and this effect is not overcome by high concentrations of Ca2+. The inhibitory effect of A23187 on the response to corticotropin, like that response itself, takes place at some part of steroid synthesis after entry of cholesterol into the cells and before the side-chain cleavage of cholesterol. These studies confirm the importance of protein synthesis in the response to corticotropin and demonstrate that the effect of protein synthesized under the influence of corticotropin is exerted at some point in the events which bring substrate (cholesterol) to the mitochondrial side-chain cleavage enzyme system. It is also shown that A23187 inhibits protein synthesis, and hence the response to corticotropin, by a mechanism which is independent of the concentration of available Ca2+.


Phosphorylation/inactivation of PTEN by Akt-independent PI3K signaling in retinal pigment epithelium.
Retinal pigment epithelium (RPE) plays a critical role in vertebrate vision by providing functional and structural support to the retina. Degeneration of RPE by cumulative oxidative stresses or acute injury frequently results in retinal degenerative diseases, notably age-related macular degeneration (AMD). Moreover, it has been shown that phosphorylation-mediated inactivation of PTEN (phosphatase and tensin homolog) in RPE is closely linked to AMD-like retinal degeneration in mice [1]. In this study, we used AMD mouse models, in which chemokine (C-C motif) ligand 2 (Ccl2) or chemokine (C-C motif) receptor 2 (Ccr2) were genetically ablated, to examine mechanisms linking reactive oxygen species (ROS) to phosphorylation/inactivation of PTEN in RPE. We found that ROS levels were increased in these RPE cells in association with phosphorylation/inactivation of PTEN. Both PTEN phosphorylation/inactivation and consequent Akt activation in the RPE of AMD model mice were inhibited by antioxidant treatment, indicating a functional role for elevated intracellular ROS. We further discovered that PTEN phosphorylation in oxidatively stressed RPE was repressed by a phosphoinositide 3-kinase (PI3K) inhibitor, but not by an Akt inhibitor. Taken together, these results suggest that ROS-activated PI3K potentiates AMD-related RPE pathogenesis through phosphorylation/inactivation of PTEN.


Intracranial meningiomas in infancy.
We report on two infants with meningioma under the age of 1 year. One is an 11-month old male infant with a fibroblastic meningioma without dural attachment to the right frontal lobe. The other is an 8-month-old male infant with a hemangiopericytic meningioma in the posterior fossa arising from the right petrous ridge. Both tumors were totally removed. Twenty-four infant meningiomas reported in the literature, including our two cases, were reviewed.


Enough is enough: Radiation doses in children with gastrojejunal tubes.
Many children with gastric feeding intolerance require postpyloric tube feeding via a gastrojejunal (GJ) tube. Placement or positioning of these tubes is typically a procedure with a low dose of radiation. Although the risk of developing cancer from radiation exposure owing to computed tomography scans is well-documented in children, the risk of cumulative radiation exposure owing to frequent GJ tube replacement often goes unnoticed in the clinical decision-making process. We sought to define the frequency and cost of GJ tube replacement, quantify the radiation doses associated with the initial placement and replacements, and assess the number of conversions to surgical jejunostomies. All pediatric patients who underwent GJ tube placement or replacement by Interventional Radiology (IR), surgery, and gastroenterology between 2010 and 2018 at a single center were reviewed. We evaluated the total cost of the initial placement and replacement of each GJ tube, the total number of replacements, and the cumulative radiation dose (mGy). We identified 203 patients who underwent GJ tube placement and/or replacement, of which 150 had radiation data available. Patients underwent a median of five GJ tube replacement procedures, and there was a wide range in the number of replacements per patient, from zero to 88. Patients were exposed to a median cumulative dose of 6.0 mGy (IQR: 2.2, 22.6). Nine percent of patients with available radiation data were exposed to more than 50 mGy, solely from GJ tube replacements. The median cost per replacement was $1170. The sum of the cost of the replacements for dislodged GJs translated to more than $1.4 million during the study period. Overall, the average dose per GJ replacement was 3.50 mGy among all patients with available data. Nine percent of patients (14/150) were exposed to greater than 50 mGy cumulative radiation solely from GJ replacements. Patients who receive more than 50 mGy of cumulative radiation dose, who undergo seven GJ tube replacements in one year, or two consecutive GJ tube replacement procedures with radiation doses exceeding 10 mGy (per replacement) should be considered for a surgical jejunostomy. IV TYPE OF STUDY: Treatment study.


Monitoring a mixed starter of Hanseniaspora vineae-Saccharomyces cerevisiae in natural must: impact on 2-phenylethyl acetate production.
The effect of simultaneous or sequential inoculation of Hanseniaspora vineae CECT 1471 and Saccharomyces cerevisiae T73 in non-sterile must on 2-phenylethyl acetate production has been examined. In both treatments tested, no significant differences in Saccharomyces yeast growth were found, whereas non-Saccharomyces yeast growth was significantly different during all days of fermentation. Independently of the type of inoculation, S. cerevisiae was the predominant species from day 3 till the end of the fermentation. The dynamics of indigenous and inoculated yeast populations showed H. vineae to be the predominant non-Saccharomyces species at the beginning of fermentation in sequentially inoculated wines, whereas the simultaneous inoculation of S. cerevisiae did not permit any non-Saccharomyces species to become predominant. Differences found in non-Saccharomyces yeast growth in both fermentations influenced the analytical profiles of final wines and specifically 2-phenylethyl acetate concentration which was two-fold increased in sequentially inoculated wines in comparison to those co-inoculated. In conclusion we have shown that H. vineae inoculated as part of a sequential mixed starter is able to compete with native yeasts present in non-sterile must and modify the wine aroma profile.


The behavior during pachynema of a normal and an inverted Y chromosome in Microtus agrestis.
The pachytene behavior of the chromosomes of Microtus agrestis (L.) (Rodentia, Arvicolidae) males carrying either the standard, or the pericentrically inverted Lund Y chromosome have been examined by electron microscopy of microspread spermatocytes. There is no synapsis between the X and either the standard or the Lund Y chromosomes during any substage of pachynema. Since synapsis is generally considered a prerequisite for crossing over, there appears to be no opportunity for crossover or chiasma formation between the X and Y in this species. The G-, C- and NOR-banded mitotic karyotypes of animals carrying the standard and Lund Y are also presented.


Neighborhood deprivation, built environment, and overweight in adolescents in the city of Oslo.
Even though the social and built environment characteristics of neighborhoods have been studied as potential determinants of social inequalities in obesity among adults, fewer studies have focused on children. Our first aim was to investigate whether there were differences in the food and physical activity environments between different neighborhood deprivation levels in the city of Oslo. We also explored whether there was an association between the prevalence of overweight (including obesity) among adolescents and (i) neighborhood deprivation levels and (ii) food and physical activity environments of the neighborhoods they live in. We conducted a food and physical activity environment mapping (using ArcGIS Pro) in all neighborhoods of Oslo, which were defined by administrative boundaries (sub-districts). The neighborhood deprivation score was calculated based on the percentage of households living in poverty, unemployment in the neighborhood, and residents with low education. A cross-sectional study including 802 seventh graders from 28 primary schools in Oslo residing in 75 out of 97 sub-districts in Oslo was also performed. MANCOVA and partial correlations were ran to compare the built environment distribution between different neighborhood deprivation levels, and multilevel logistic regression analyses were used to explore the effect of neighborhood deprivation and the food and physical activity environments on childhood overweight. We found that deprived neighborhoods had greater availability of fast food restaurants and fewer indoor recreational facilities compared to low-deprived neighborhoods. Additionally, we observed that the residential neighborhoods of the adolescents with overweight had greater availability of grocery and convenience stores when compared to the residential neighborhoods of the adolescents without overweight. Adolescents living in neighborhoods with high deprivation had a two-fold higher odds (95% CI = 1.1-3.8) to have overweight compared to adolescents living in neighborhoods with low deprivation, regardless of participants' ethnicity and parental education. However, the built environment did not determine the relationship between neighborhood deprivation and overweight in adolescents. The neighborhoods in Oslo with higher deprivation levels had more obesogenic characteristics than the low-deprived neighborhoods. Adolescents living in high-deprived neighborhoods were more likely to have overweight than their counterparts from low-deprived neighborhoods. Thus, preventive measures targeting adolescents from high-deprived neighborhoods should be put in place in order to reduce incidence of overweight.


[Alveolar microlithiasis in children. Contribution of bronchoalveolar lavage].
This case of pulmonary alveolar microlithiasis emphasizes the rarity of the disease and its exceptional diagnosis in infants which relies on the pulmonary pathologic study. Besides allowing for diagnosis, the study of the alveolar brushing fluid permitted to describe for the first time pictures of post-necrotic cellular calcification which is probably responsible for the disease.


Association between fetal vascular malperfusion and gestational diabetes.
Diabetes mellitus (DM) is a major complication in pregnancy. Placental lesions with DM remain unclear and controversial. Recently, the terms of placental pathological findings, such as maternal and fetal vascular malperfusions (MVM and FVM, respectively) were introduced by the Amsterdam Placental Workshop Group Consensus Statement (APWGCS). FVM cases were classified as the partial obstruction type (global FVM) and the complete obstruction type (segmental FVM). The aim of this study was to clarify the pathological characteristics of the placenta with pregestational DM/gestational DM; GDM according to APWGCS. We studied the placentas of 182 DM women (27 pregestational DM and 155 GDM) and control placentas of 460 women without DM during 2011-2018. We excluded cases of intrauterine fetal death or multiple pregnancies. We reviewed microscopical findings including, MVM, FVM, chorioamnionitis with the slides according to the APWGCS. Microscopically, the incidence of FVM was significantly higher in GDM patients than control (17% vs. 10%, p = 0.0138), but not significant in pregestational DM (11%, p = 0.7410). Segmental FVM (complete obstruction) was significantly more observed in GDM than control group (5% vs. 0.4%, p = 0.0013). Segmental FVM in GDM showed high incidence of light-for-dates infant (three of seven cases, 43%, p = 0.0288). In addition, several segmental FVM findings (villous stromal-vascular karyorrhexis and stem vessel occlusion) were frequently noted in 2 or 3 points positive of 75 g oral glucose tolerance test than 1 point positive GDM. Our placental findings suggest disorder of carbohydrate metabolism might affect the fetal vascular damage, especially complete fetal vascular obstruction.


Pathological angiogenesis and inflammation in tissues.
The role of angiogenesis in the growth of organs and tumors is widely recognized. Vascular-organ interaction is a key mechanism and a concept that enables an understanding of all biological phenomena and normal physiology that is essential for human survival under pathological conditions. Recently, vascular endothelial cells have been classified as a type of innate immune cells that are dependent on the pathological situations. Moreover, inflammatory cytokines and signaling regulators activated upon exposure to infection or various stresses play crucial roles in the pathological function of parenchymal cells, peripheral immune cells, stromal cells, and cancer cells in tissues. Therefore, vascular-organ interactions as a vascular microenvironment or tissue microenvironment under physiological and pathological conditions are gaining popularity as an interesting research topic. Here, we review vascular contribution as a major factor in microenvironment homeostasis in the pathogenesis of normal as well as cancerous tissues. Furthermore, we suggest that the normalization strategy of pathological angiogenesis could be a promising therapeutic target for various diseases, including cancer.


Uric acid demonstrates neuroprotective effect on Parkinson's disease mice through Nrf2-ARE signaling pathway.
Uric acid has neuroprotective effect on Parkinson's disease (PD) by inhibiting oxidative damage and neuronal cell death. Our previous study has shown that uric acid protected dopaminergic cell line damage through inhibiting accumulation of NF-E2-related factor 2 (Nrf2). This study aimed to investigate its in vivo neuroprotective effect. PD was induced by MPTP intraperitoneally injection for 7 d in male C57BL/6 mice. Mice were treated with either uric acid (intraperitoneally injection 250 mg/kg) or saline for a total of 13 d. We showed that uric acid improved behavioral performances and cognition of PD mice, increased TH-positive dopaminergic neurons and decreased GFAP-positive astrocytes in substantia nigra (SN). Uric acid increased mRNA and protein expressions of Nrf2 and three Nrf2-responsive genes, including γ-glutamate-cysteine ligase catalytic subunit (γ-GCLC), heme oxygenase-1 (HO-1) and NQO1. Uric acid significantly increased superoxide dismutase (SOD), CAT, glutathione (GSH) levels and decreased malondialdehyde (MDA) level in SN regions of MPTP-treated mice. Uric acid inhibited the hippocampal expression of IL-1β and decreased serum and hippocampus levels of interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α). In conclusion, uric acid demonstrates neuroprotective properties for dopaminergic neurons in PD mice through modulation of neuroinflammation and oxidative stress.


Transient and multivariate system for transformation of a fungal plant pathogen, Rosellinia necatrix, using autonomously replicating vectors.
Rosellinia necatrix is a fungus that infects a wide range of host plants and ruins a variety of commercially important crops. DNA fragments can be introduced into R. necatrix using conventional protoplast-PEG transformation and genome-integrating vectors; however, transformation efficiency with this strategy is quite low. Therefore, to establish a more effective transformation system for the studies of R. necatrix, an autonomously replicating vector was constructed using AMA1 sequences derived from Aspergillus nidulans, which is distantly related to R. necatrix. Use of this vector with AMA1 sequences increased transformation efficiency in R. necatrix, and the vector was maintained as a plasmid in the transformants. Transient and multivariate functional analyses in R. necatrix were performed using co-transformation of multiple pAMA-H vectors, which each carried either an expression cassette for eGFP, mOrange2, or a geneticin resistance gene. Furthermore, fluorescent proteins expressed from the autonomously replicating vectors were dispersed throughout fungal colonies even though the vectors themselves were restricted to the center of each colony. This intriguing phenomenon indicated that gene products could move from the center to the margin in a colony of the filamentous fungi via a cell-to-cell transport system.


Minimal-preparation abdomino-pelvic CT in frail and elderly patients: prognostic value of colonic and extracolonic findings.
To examine the overall survival of patients who had had been referred for minimal preparation abdomino-pelvic computed tomography (MPCT), and to assess the prognostic value of the colonic and extracolonic findings detected. The survival of a cohort of 1029 elderly and frail patients, with clinical symptoms and signs suspicious for colorectal cancer (CRC), who had undergone MPCT between 1995 and 1998 was investigated. Univariate and multivariate survival analyses were undertaken according to the presence of CRC and extracolonic abnormalities (ECA). The median age of the 1029 patients was 79.4 years. The overall median survival following MPCT was 5.4 years; and 6.6 years if no abnormality was detected. On multivariate analysis, age, sex, CRC status, and number of ECAs were significant factors in overall survival. Median survival for those with confirmed CRC [n=91 (prevalence, 8.8%)] was 1.1 years, compared with 5.9 years without CRC (p<0.0001); and 2.4 years for those with one or more ECA [n=245 (prevalence, 23.8%)], compared with 6.1 years without ECA (p<0.0001). Survival was progressively shorter for increasing numbers of ECAs; and shorter for previously unknown non-CRC malignancies (n=24) compared with CRC (p<0.0001). MPCT appears to have prognostic potential in this patient population, with significant reductions in survival if a CRC or ECA is detected. The detection of ECA would appear to have at least as important an impact on the usefulness of the examination as the detection of CRC.


Bifid ureter with blind-ending branch.
A patient with a blind-ending branch of bifid ureter, one of the rarest anomalies in the upper urinary tract, is described.


Fenfluramine binds 5-hydroxytryptophan.
Fenfluramine, an anorexigenic drug, lowers serotonin (5-hydroxytryptamine) and 5-hydroxyindoleacetic acid levels in brain, spinal fluid, and blood, and has been used as a treatment for autism. Fenfluramine's mode of action is unknown. We present evidence from chromatography and nuclear magnetic resonance spectroscopy that fenfluramine selectively binds the serotonin and 5-hydroxyindoleacetic acid precursor, 5-hydroxytryptophan. The mode of binding may have general applications for the understanding of drug activity, receptor binding, and for the design of specific antagonists to aromatic compounds.


Human teratocarcinoma stem cells: glycolipid antigen expression and modulation during differentiation.
Teratocarcinomas are germ cell tumors in which pluripotent stem cells, embryonal carcinoma (EC) cells, undergo differentiation along the pathways resembling those occurring during early embryogenesis. Human EC cell lines established in vitro provide a model for studying embryonic cellular differentiation in a way that is pertinent to early human development. The predominant glycolipid antigens expressed by EC cells of both humans and mice have globoseries core structures; in humans they are terminally modified to yield the monoclonal antibody-defined stage-specific embryonic antigens SSEA-3 and SSEA-4, and also globo-ABH antigens; in the mouse terminal modification yields the Forssman antigen rather than SSEA-3 and -4. These observations focus attention on the possible role of the P-blood group system, which regulates synthesis of globoseries oligosaccharides, in the behavior of cells in the early embryo and in teratocarcinomas. Marked changes in the core structures of the cell surface glycolipids occur as the EC cells differentiate; thus globoseries structures rapidly diminish and are replaced by lactoseries and then by ganglioseries glycolipids. During differentiation of the NTERA-2 line of pluripotent human EC cells into neurons and other cell types, the various subsets of differentiated cells that arise are distinguished by their differential expression of new glycolipid antigens, particularly ganglioside GT3 (recognized by antibody A2B5), and ganglioside 9-0-acetyl GD3 (recognized by antibody ME311). Neurons are found among the A2B5+/ME311- cells.


Burden of community-associated Clostridioides difficile infection in southeastern United States: a population-based study.
This cross-sectional population-based study aims to determine overall incidence rate of Clostridioides difficile infection (CDI) in the State of South Carolina and provide an estimated cost of hospitalization due to community-associated CDI (CA-CDI). All CDI cases in South Carolina were identified through National Healthcare Safety Network (NHSN) and the South Carolina Infectious Disease and Outbreak Network (SCION) from January 1, 2015 to June 30, 2016, excluding infants < 1 year of age. During the 18-month study period, 10,254 CDI events were identified in South Carolina residents with an overall incidence rate of 139/100,000 person-years. Over one-half of CDI cases were CA-CDI (5192; 51%) with an incidence rate of 71/100,000 person-years. Among patients with CA-CDI, 2127 (41%) required hospitalization with a median length of stay of 5 days. The annual burden of CA-CDI in South Carolina was estimated to be 9282 hospital days and $16,217,295 in hospitalization costs. The incidence rate of CA-CDI in South Carolina has surpassed both community-onset healthcare facility associated and hospital-onset CDI combined. The heavy burden of CA-CDI justifies dedication of public health resources to combat CDI in ambulatory settings, through antimicrobial stewardship initiatives.


Enhanced Stim1 expression is associated with acquired chemo-resistance of cisplatin in osteosarcoma cells.
Osteosarcoma is the most common primary malignant bone tumor. Although cisplatin is the primary chemotherapy used in osteosarcoma treatment, the cisplatin resistance remains a big challenge for improving overall survival. The store-operated calcium (Ca2+) entry (SOCE) and its major mediator Stim1 have been shown to be implicated in a number of pathological processes typical for cancer. In this study, we showed that Stim1 expression was significantly increased in chemo-resistant osteosarcoma tissues compared with chemo-sensitivity tissues. Patients with Sitm1 expression exhibited poorer overall survival than Stim1-negative patients. Moreover, un-regulation of Stim1 expression and SOCE were also observed in cisplatin-resistant MG63/CDDP cells compared with their parental cells. Cisplatin treatment obviously reduced Stim1 expression and SOCE in cisplatin-sensitivity MG63 cells, but had no effects on MG63/CDDP cells. In addition, cisplatin resulted in a more pronounced increase of endoplasmic reticulum (ER) stress in MG63 cells than in their resistant variants, which was evidenced by the activation of molecular markers of ER stress, GRP78, CHOP and ATF4. Knockdown of Stim1 using siRNA remarkably enhanced cisplatin-induced apoptosis and ER stress in MG63/CDDP cells, thereby sensitizing cancer cells to cisplatin. On the other hand, overexpression of Stim1 markedly reversed apoptosis and ER stress following cisplatin treatment. Taken together, these results demonstrate that Stim1 as well as Ca2+ entry contributes cisplatin resistance via inhibition of ER stress-mediated apoptosis, and provide important clues to the mechanisms involved in cisplatin resistance for osteosarcoma treatment. Stim1 represents as a target of cisplatin and blockade of Stim1-mediated Ca2+ entry may be a useful strategy to improve the efficacy of cisplatin to treat osteosarcoma.


Professional identity and its influencing factors of first-year post-associate degree baccalaureate nursing students: A cross-sectional study.
Professional identity is an important and universal concept in the field of nursing because it not only affects nurses' perceptions of their role in nursing, but it also affects retention rates. However, the influential factors that impact the professional identity of nursing students currently are not well known. This exploratory study aims to investigate the concept of professional identity and confirm its influencing factors among post-associate degree baccalaureate nursing students in China. We conducted a cross-sectional study of 198 first-year post-associate degree baccalaureate nursing students enrolled in full-time study at two provincial medical colleges in China. We used paper and pencil questionnaires to conduct the survey and obtain the results used in this study. The overall mean score for 'professional identity' of the sample was 3.63 ± 0.62 on a scale of 1 through 5, with the means for the five dimensions of professional identity ranging from 3.29 to 4.02. A clinical learning experience that is longer than eight months (ref. = 8 months) (β = 0.138, p < 0.036) and a positive perception of the clinical learning environment (β = 0.476, p < 0.001) are the dominant factors that we found to be positively associated with professional identity. Nurse educators should investigate the factors that influence professional identity among their students to help develop a stable and satisfied nursing workforce.


Incidence, risk factors and economic impact of clinical mastitis in dromedary camel (Camelus dromedarius).
The study aimed to investigate the incidence, risk factors and economic impact of clinical mastitis in an organized camel herd located at Bikaner, Rajasthan, India. Furthermore, bacterial pathogens were isolated and identified using MALDI-TOF mass spectrometry analysis. The overall annual incidence of clinical mastitis was 25%, with highest incidence in Bikaneri and Mewari breed followed by Kachchhi and Jaisalmeri breeds. Animals with high peak yield appeared at higher risk, though statistical significance could not be established for any risk factor included in the study. Highest incidence was recorded in camels falling in 4th or higher parity group. In 80% cases, only one udder quarter was involved, while in rest 20% cases, two udder quarters were involved. Quarter-wise incidence was maximum in right fore-quarter followed by left fore-quarter, right hind-quarter and left hind-quarter. Important pathogens identified included Staphylococcus aureus, E. coli, Klebsiella pneumoniae, Staphylococcus epidermidis, Streptococcus uberis, Bacillus clausii and Corynebacterium amylocolatum. Other isolates included Staphylococcus hominis, Streptococcus pluranimalium, Aerococcus viridians and Moraxella osloensis. The mean cost of clinical mastitis per animal per month was calculated as Indian Rs. 181.63 (~ US$ 2.45), while the mean cost per animal per annum was Indian Rs. 2179.29 (~ US$ 29.37). The maximum cost incurred in purchase of drugs (43.63%), followed by cost of discarded milk plus milk production loss (18.17%) and cost of veterinary services (17.03%). To our knowledge, this is the first report documenting economic impact of clinical mastitis in camel.


Tonic activity of Galpha-gustducin regulates taste cell responsivity.
The taste-selective G protein, alpha-gustducin (alpha-gus) is homologous to alpha-transducin and activates phosphodiesterase (PDE) in vitro. alpha-Gus-knockout mice are compromized to bitter, sweet and umami taste stimuli, suggesting a central role in taste transduction. Here, we suggest a different role for Galpha-gus. In taste buds of alpha-gus-knockout mice, basal (unstimulated) cAMP levels are high compared to those of wild-type mice. Further, H-89, a cAMP-dependent protein kinase inhibitor, dramatically unmasks responses to the bitter tastant denatonium in gus-lineage cells of knockout mice. We propose that an important role of alpha-gus is to maintain cAMP levels tonically low to ensure adequate Ca2+ signaling.


Contemporary Data on the Status and Medical Management of Acute Heart Failure.
Acute heart failure (AHF) is among the leading causes for unplanned hospital admission. Despite advancements in the management of chronic heart failure, the prognosis of AHF remains poor with high in-hospital mortality and increased rates of unfavorable post-discharge outcomes. With this review, we aim to summarize current data on AHF epidemiology, focus on the different patient profiles and classifications, and discuss management, including novel therapeutic options in this area. There is significant heterogeneity among patients admitted for AHF in their baseline characteristics, heart failure (HF) aetiology and precipitating factors leading to decompensation. A novel classification scheme based on four distinct clinical scenarios has been included in the most recent ESC guidelines, in an effort to better risk stratify patients and guide treatment. Intravenous diuretics, vasodilators, and inotropes remain the cornerstone of management in the acute phase, and expansion of use of mechanical circulatory support has been noted in recent years. Meanwhile, many treatments that have proved their value in chronic heart failure demonstrate promising results in the setting of AHF and research in this field is currently ongoing. Acute heart failure remains a major health challenge with high in-hospital mortality and unfavorable post-discharge outcomes. Admission for acute HF represents a window of opportunity for patients to initiate appropriate treatment as soon as possible after stabilization. Future studies are needed to elucidate which patients will benefit the most by available therapies and define the optimal timing for treatment implementation.