Prognostic relevance of papillary muscle infarction in reperfused infarction as visualized by cardiovascular magnetic resonance.
The prognostic significance of papillary muscle infarction (PapMI) on hard clinical outcomes has not been investigated in patients with reperfused ST-segment-elevation myocardial infarction. Noninvasive investigation by gadolinium-enhanced cardiac MRI enables the detection of PapMI with high spatial resolution. The aim of our study was (1) to assess the incidence, determinants, and clinical characteristics of PapMI in a large multicenter cohort of patients with ST-segment-elevation myocardial infarction and (2) to assess the prognostic significance of PapMI at 1-year follow-up. We enrolled 738 patients with ST-segment-elevation myocardial infarction reperfused by primary angioplasty (<12 hours after symptom onset) in this cardiac MRI study at 8 centers. Cardiac MRI was completed within 1 week after infarction using a standardized protocol. Central core laboratory-masked analyses for the presence of PapMI were performed. The primary clinical end point of the study was the occurrence of major adverse cardiac events. PapMIs were detected in 104 patients (14%). The presence of PapMI was associated with larger infarcts (P<0.001), less myocardial salvage (P<0.001), impaired left ventricular function (P<0.001), and more pronounced reperfusion injury (P=0.02). Patients with PapMI had a significantly higher mortality (8 [7.7%] versus 12 [1.9%]) and major adverse cardiac events (21 [20.2%] versus 31 [4.9%]) rate at 12-month follow-up (P<0.001, respectively). PapMI was identified as a significant independent predictor of major adverse cardiac events (hazard ratio, 4.41 [confidence interval, 2.54-7.68]; P<0.001). The presence of PapMI is associated with decreased myocardial salvage, larger infarcts, and more pronounced reperfusion injury with subsequent significantly increased major adverse cardiac event rates. Consequently, our data underscore the importance of PapMI as a marker of poor outcome in patients with ST-segment-elevation myocardial infarction.


TGF-beta 1 inhibition of transin/stromelysin gene expression is mediated through a Fos binding sequence.
Transforming growth factor beta 1 (TGF-beta 1) inhibits the growth factor and oncogene induction of transin/stromelysin, a secreted matrix-degrading metalloprotease. We demonstrate that a 10 bp element in the transin promoter is required for the TGF-beta 1 inhibitory effects and that this sequence is conserved in the promoter regions of several other TGF-beta 1-inhibited genes. The TGF-beta 1 inhibitory element (TIE) specifically binds a nuclear protein complex from TGF-beta 1-stimulated rat fibroblasts. Interestingly, this complex contained the c-fos proto-oncogene product, Fos, and induction of Fos expression was required for the inhibitory effect of TGF-beta 1 on transin gene expression. These results suggest that TGF-beta 1 inhibition of gene expression is mediated by the binding of a Fos-containing protein complex to the TIE promoter sequences.


[Sleepiness and accidental risk].
Sleepiness while driving is both a very common phenomena regarding 10 to 15% of drivers and one of the main causes of death on highways (one third of fatal accidents). Sleepiness results mainly from behavioural causes: sleep deficit or irregular work schedules. However, many sleep diseases and hypnotics may also induce sleepiness at the wheel. Several objective and subjective measures have been validated to assess sleepiness. Road safety and information to the drivers have now to focus on better informing drivers on the rules of preventing sleepiness at the wheel in order to avoiding hundred of accidents in the next future.


An echocardiographic study of tetralogy of Fallot in the fetus and infant.
To document the echocardiographic features of tetralogy of Fallot during fetal and postnatal life. Correlation of echocardiographic findings with the requirement for early intervention prior to definitive repair. Retrospective observational study. A tertiary fetal cardiology unit. Fetuses with a diagnosis of tetralogy of Fallot identified from a prospective database between 1 January 1999 and 31 October 2002. MAIN MEASURES OF OUTCOME: Growth of aorta and pulmonary trunk during fetal and postnatal life. Doppler assessment of the great arteries both prenatally and postnatally. Clinical outcome to definitive repair. We identified 25 fetuses with tetralogy of Fallot, 23 having a pulmonary valvar diameter below the normal range at some point during gestation. The ratio of the diameter of the aortic to the pulmonary valve was abnormal in all cases. The pulmonary arterial Doppler velocity was within the normal range in six fetuses at presentation,and elevated in the remainder. In two fetuses, the right ventricular outflow tract was patent during fetal life, but had become atretic at birth. Both of these fetuses had reversal of flow in the arterial duct at presentation during fetal life. In 2 fetuses in whom we showed poor growth of the pulmonary trunk in late gestation, it was necessary to intervene early. The Doppler velocity across the pulmonary valve during fetal life did not differentiate between babies who required early intervention and those who were repaired electively. There was a marked increase in pulmonary arterial Doppler velocity following birth, which became more elevated with age. Of 18 liveborn infants, 17 have survived, with 2 having balloon dilation of the right ventricular outflow tract, and 3 insertion of a Blalock-Taussig shunt prior to definitive repair. In tetralogy of Fallot, features of pulmonary valvar hypoplasia and obstruction are evident during fetal life. Progression of obstruction in the right ventricular outflow tract was observed during fetal life as well as postnatally. Reversal of flow in the arterial duct, and failure of growth of the pulmonary trunk, predicted the need for early surgery to maintain pulmonary blood flow. Parents should be counselled about the possibility of emergency intervention being required after birth. Affected fetuses should be delivered at units with experience of managing the cyanosed newborn.


Does Aseptic Revision Risk Differ for Primary Total Knee Arthroplasty Patients Who Have and Do not Have a Prior Primary or Revision Arthroplasty?
We sought to evaluate the risk of aseptic revision in total knee arthroplasty (TKA) patients who have and do not have a history of primary or revision arthroplasty of a different major joint. We conducted a matched cohort study using data from Kaiser Permanente's arthroplasty registries. Patients who underwent primary unilateral TKA (index knee) were identified (2009-2018). Two matches based on exposure history were performed: (1) 33,714 TKAs with a history of primary arthroplasty of a different joint (contralateral knee, either hip, and/or either shoulder) were matched to 67,121 TKAs without an arthroplasty history and (2) 597 TKAs with a history of aseptic revision in a different joint were matched to 1,190 TKAs with a history of a prior arthroplasty in a different joint, but without any revision. After the matches were performed, Cox regressions were used to evaluate aseptic revision risk of the index knee using the no history groups as the reference in regression models. No difference in aseptic revision risk for the index knee was observed when comparing patients who had a prior primary arthroplasty in a different joint to those who did not have an arthroplasty history (hazard ratio = 0.95, 95% CI = 0.86-1.06). Those patients who did not have any prior aseptic revision history in a different joint had higher risk of aseptic revision in the index knee (hazard ratio = 2.06, 95% CI = 1.17-3.63). Patients who had a prior revision history had over a 2-fold higher risk of aseptic revision in the index knee, warranting close surveillance of these patients. Level III.


Highly selective CO2 removal for one-step liquefied natural gas processing by physisorbents.
Industrial specifications require CO2 concentrations in natural gas below 50 ppm during liquefaction because of corrosion and CO2 freezing. Herein, we report a physisorbent (TIFSIX-3-Ni) that exhibits new benchmark CO2/CH4 selectivity and fast kinetics, thereby enabling one-step LNG processing to CO2 levels of 25 ppm.


Cell density-dependent expression of chondroitin sulfate proteoglycan in cultured human monocytes.
Monocytes were isolated and established in vitro at different cell densities. The incorporation of [35S]sulfate into macromolecules in monocytes (day 1 in culture) and monocyte-derived macrophages (day 5 in culture) was found to increase with decreasing cell density in approximately the same way in both day 1 and day 5 cell cultures. [35S]Sulfate was found to be incorporated almost exclusively into chondroitin sulfate proteoglycan (CSPG) in both high and low density monocyte and monocyte-derived macrophage cultures. The molecular size of the [35S]CSPGs produced by the high and low cell density cultures were not found to differ as judged by gel chromatography elution patterns. The molecular size and the structure of the glycosaminoglycan chains were found to be almost similar in high and low density day 1 and day 5 cultures. Only a small degree of proteoglycan degradation could be observed in both high and low density cultures. Furthermore, cell density-dependent differences in CSPG biosynthesis could be observed already 2 h after the establishment of the cultures, indicating that a process of "down-regulation" in high density cultures was already in operation. The glycosaminoglycan synthesis in high cell density day 1 cultures could be increased slightly following exposure to 0.5 mM benzyl-beta-D-xyloside, but not to the same level as that observed in untreated low cell density cultures. By contrast, the expression of 35S-macromolecules by cells cultured at high cell density for 5 days could be increased by xyloside treatment almost to the same level as that observed in the low density cultures.


Severity of GNAO1-Related Disorder Correlates with Changes in G-Protein Function.
GNAO1-related disorders (OMIM #615473 and #617493), caused by variants in the GNAO1 gene, are characterized by developmental delay or intellectual disability, hypotonia, movement disorders, and epilepsy. Neither a genotype-phenotype correlation nor a clear severity score have been established for this disorder. The objective of this prospective and retrospective observational study was to develop a severity score for GNAO1-related disorders, and to delineate the correlation between the underlying molecular mechanisms and clinical severity. A total of 16 individuals with GNAO1-related disorders harboring 12 distinct missense variants, including four novel variants (p.K46R, p.T48I, p.R209P, and p.L235P), were examined with repeated clinical assessments, video-electroencephalogram monitoring, and brain magnetic resonance imaging. The molecular pathology of each variant was delineated using a molecular deconvoluting platform. The patients displayed a wide variability in the severity of their symptoms. This heterogeneity was well represented in the GNAO1-related disorders severity score, with a broad range of results. Patients with the same variant had comparable severity scores, indicating that differences in disease profiles are not due to interpatient variability, but rather, to unique disease mechanisms. Moreover, we found a significant correlation between clinical severity scores and molecular mechanisms. The clinical score proposed here provides further insight into the correlation between pathophysiology and phenotypic severity in GNAO1-related disorders. We found that each variant has a unique profile of clinical phenotypes and pathological molecular mechanisms. These findings will contribute to better understanding GNAO1-related disorders. Additionally, the severity score will facilitate standardization of patients categorization and assessment of response to therapies in development. ANN NEUROL 2023;94:987-1004.


Correction: The Role of Aerobic Fitness in Cortical Thickness and Mathematics Achievement in Preadolescent Children.
[This corrects the article DOI: 10.1371/journal.pone.0134115.].


Ethanol induces volume changes and gap junction closure via intracellular Ca2+ signalling pathway in cultured rabbit gastric epithelial cells.
Ethanol is a well-established 'barrier breaker' in gastric mucosa, but its effects at cellular level remain to be detailed. Gastric epithelial cells were isolated from rabbits and cultured to monolayers. Intracellular calcium was measured spectrofluorometrically with fura-2. The patency of gap junctions was assessed by photobleaching a small area of 5-carboxyfluorescein loaded monolayer and measuring recovery of fluorescence. For cell volume measurements the change in fluorescence intensity was followed in calcein-loaded monolayers with a confocal microscope. Intracellular calcium concentration was increased from 65 +/- 9 to 140 +/- 17 nM; recovery of fluorescence signal after photobleaching was diminished from 53% +/- 11% to 9% +/- 3%; and cell volume was decreased significantly after 10 min exposure to 5% (vol/vol) ethanol. This volume decrease was prevented with serosal application of the potassium channel blocker, quinine, or by blocking the intracellular calcium signalling pathway with the intracellular calcium-chelating agent BAPTA. This suggests that luminal ethanol opens the basolateral calcium-dependent potassium selective channels via calcium signalling pathway, with resultant shrinkage of the cell. Intracellular calcium concentration is increased, gap junctions are closed and cell volume is decreased after exposure to 5% ethanol. Since gap junctions are known to be calcium gated, it is likely that their closure is secondary to the elevated cytosolic calcium in ethanol injured cells. This may have a protective function by limiting intercellular spread of impending cell injury. The opening of the basolateral potassium channel probably underlies the ethanol-induced cell shrinkage and might contribute to the ethanol-provoked epithelial damage.


The Anatomy of the Frontal Process of the Maxilla in the Medial Wall of the Lacrimal Drainage System in East Asians.
To investigate the anatomical structure characteristics of the frontal process of the maxilla in the medial wall of the lacrimal drainage system for endoscopic dacryocystorhinostomy based on the cone-beam computed tomography images. One hundred eight sides of orbits were retrospectively evaluated in this study. The angulation and thickness of the frontal process of the maxilla in the medial wall of the lacrimal drainage system were measured and compared at 2 different levels: lacrimal sac level and nasolacrimal duct level. The vertical height between the 2 measurement levels was also measured. Data were compared between males and females. The average angulation and thickness of the frontal process of the maxilla were found significantly larger at the lacrimal sac level than at the nasolacrimal duct level (83.1 ± 11.3° vs. 61.7 ± 10.9° and 2.7 ± 0.9 mm vs. 1.1 ± 0.4 mm) (p < 0.001). The vertical height between the 2 levels was significantly higher in males than in females (8.6 ± 1.8 mm vs. 7.8 ± 1.7 mm) (p = 0.015). We found the angulation and thickness of the frontal process of the maxilla in the medial wall of the lacrimal drainage system decreased from the upper to the lower level. Starting the osteotomy during endoscopic dacryocystorhinostomy might be easier and safer at the midpoint level of the maxillary line.


Thin-layer chromatography of chlorophylls and their derivatives on cellulose layers.
A method for the separation and identification of chlorophyll a and b, pheophytin a and b, pheophorbide a and b, chlorin e6, rhodin g7, and the corresponding chlorophyll a' and b' derivatives, as well as the saponification products of pheophorbide a and b, by thin-layer chromatography on commercial cellulose layers on the micro-scale has been developed. Two solvent systems were used: light petroleum (b.p.60-80 degrees)-pyridine (9:1, v/v) and n-heptane-pyridine (7:3, v/v). The former was suitable for chlorophylls, pheophytins and pheophorbides, and the latter for pheophorbides, chlorin, rhodin and their esters. The separation of the derivatives was good and no chemical alteration of the derivatives could be observed. The method is rapid and easy to use and is therefore suitable for checking the purity of derivatives during the preparation of chlorophylls and their derivatives.


CAESAR: a new tool to assess relatives' experience of dying and death in the ICU.
To develop an instrument designed specifically to assess the experience of relatives of patients who die in the intensive care unit (ICU). The instrument was developed using a mixed methodology and validated in a prospective multicentre study. Relatives of patients who died in 41 ICUs completed the questionnaire by telephone 21 days after the death, then completed the Hospital Anxiety and Depression Scale, Impact of Event Scale-Revised and Inventory of Complicated Grief after 3, 6, and 12 months. A total of 600 relatives were included, 475 in the main cohort and 125 in the reliability cohort. The 15-item questionnaire, named CAESAR, covered the patient's preferences and values, interactions with/around the patient and family satisfaction. We defined three groups based on CAESAR score tertiles: lowest (≤59, n = 107, 25.9 %), middle (n = 185, 44.8 %) and highest (≥69, n = 121, 29.3 %). Factorial analysis showed a single dimension. Cronbach's alpha in the main and reliability cohorts was 0.88 (0.85-0.90) and 0.85 (0.79-0.89), respectively. Compared to a high CAESAR score, a low CAESAR score was associated with greater risks of anxiety and depression at 3 months [1.29 (1.13-1.46), p = 0.001], post-traumatic stress-related symptoms at 3 [1.34 (1.17-1.53), p < 0.001], 6 [OR = 1.24 (1.06-1.44), p = 0.008] and 12 [OR = 1.26 (1.06-1.50), p = 0.01] months and complicated grief at 6 [OR = 1.40 (1.20-1.63), p < 0.001] and 12 months [OR = 1.27 (1.06-1.52), p = 0.01]. The CAESAR score 21 days after death in the ICU is strongly associated with post-ICU burden in the bereaved relatives. The CAESAR score should prove a useful primary endpoint in trials of interventions to improve relatives' well-being.


Polybrominated diphenyl ethers (PBDEs) and alternative brominated flame retardants (aBFRs) in sediments from four bays of the Yellow Sea, North China.
The distribution characteristics and potential sources of polybrominated diphenyl ethers (PBDEs) and alternative brominated flame retardants (aBFRs) were investigated in 54 surface sediment samples from four bays (Taozi Bay, Sishili Bay, Dalian Bay, and Jiaozhou Bay) of North China's Yellow Sea. Of the 54 samples studied, 51 were collected from within the four bays and 3 were from rivers emptying into Jiaozhou Bay. Decabromodiphenylethane (DBDPE) was the predominant flame retardant found, and concentration ranged from 0.16 to 39.7 ng g(-1) dw and 1.13-49.9 ng g(-1) dw in coastal and riverine sediments, respectively; these levels were followed by those of BDE 209, and its concentrations ranged from n.d. to 10.2 ng g(-1) dw and 0.05-7.82 ng g(-1) dw in coastal and riverine sediments, respectively. The levels of DBDPE exceeded those of decabromodiphenyl ether (BDE 209) in most of the samples in the study region, whereas the ratio of DBDPE/BDE 209 varied among the four bays. This is indicative of different usage patterns of brominated flame retardants (BFRs) and also different hydrodynamic conditions among these bay areas. The spatial distribution and composition profile analysis indicated that BFRs in Jiaozhou Bay and Dalian Bay were mainly from local sources, whereas transport from Laizhou Bay by coastal currents was the major source of BFRs in Taozi Bay and Sishili Bay. Both the ∑PBDEs and ∑aBFRs (sum of pentabromotoluene (PBT), 2,3-diphenylpropyl-2,4,6-tribromophenyl ether (DPTE), pentabromoethylbenzene (PBEB), and hexabromobenzene (HBB)) were at low concentrations in all the sediments. This is probably attributable to a combination of factors such as low regional usage of these products, atmospheric deposition patterns, coastal currents transportation patterns, and degradation processes for higher BDE congeners. This paper is the first study that has investigated the levels of DBDPE in the coastal sediments of China's Yellow Sea.


High-efficiency gene transfer into adult fish: a new tool to study fin regeneration.
Zebrafish represents an excellent model to study the function of vertebrate genes (e.g., well-developed genetics, large number of mutants, and genomic sequencing in progress), inasmuch as we have tools to manipulate gene expression. Recent use of injected morpholinos in eggs provides a good method to " knockdown " gene expression in early development (Nasevicius and Ekker, 2000), and the "caged" RNA injected in eggs allows to overexpress a gene in a specific set of cells (Ando et al., 2001). However, a method to specifically modify gene expression in the juvenile or in the adult is still missing. Such a method would be a very powerful tool to understand gene function in differentiated tissues. We describe here an electroporation-based approach, which allows gene transfer in adult tissues. Its efficiency was assessed using a GFP (green fluorescent protein) dependent assay. We then used this method to disrupt the Fgf signalling pathway during the process of regeneration.


Natural History and Outcomes of Cavernous Transformation of the Portal Vein in Cirrhosis.
Cavernous transformation of the portal vein can occur after portal vein thrombosis (PVT). In this study, we investigated clinical complications associated with cavernous transformation in the context of cirrhosis and PVT. In this retrospective cohort analysis, 204 patients with cirrhosis and PVT with or without cavernous transformation were identified using MUSC's Clinical Data Warehouse between January 1, 2013, through December 31, 2019. Complete demographic data, clinical history, and laboratory tests were abstracted from the electronic medical record. Of 204 patients, 41 (20%) had cavernous transformation. MELD, Child-Pugh, and Charlson Comorbidity Index scores were similar among groups. There were no significant differences in the prevalence of esophageal varices (with or without bleeding), splenomegaly, or hepatic encephalopathy in patients with and without cavernous transformation, although ascites tended to be lower in patients with cavernous transformation (31/41 (76%) vs 142/163 (87%), p = 0.06). Patients with cavernous transformation were significantly less likely to have hepatocellular carcinoma (HCC) (13/41 (32%) vs 81/163 (50%), p < 0.05) and had significantly lower APRI (1.4 vs 2.0, p < 0.05) and Fib-4 (4.7 vs 6.5, p < 0.05). Patients with cavernous transformation had lower 5-year mortality (12/41 (29%) vs 81/163 (49%) died, p = 0.06). The 10-year mortality of patients with cavernous transformation without HCC was significantly lower than in those without cavernous transformation (8/28 (29%) vs 46/82 (56%), respectively, p < 0.05). Patients with cavernous transformation appeared to have better outcomes than those without cavernous transformation.


Variability in growth and reproduction in F1-families of an Erophila verna population.
Seeds from two morphologically different groups in an Erophila verna dune population were used in a growth experiment to analyse and quantify flexibility and plasticity of growth and reproductive characteristics.A strong correlation between plant morphology and individual seed weight was shown in parents and progeny. Seeds of plants with narrow leaves were twice as heavy (H-plants) as those from plants with broad leaves (L-plants) Families of H-plants allocated ca. 10% more dry matter to the roots. These characters were fixed, presumably due to selfing. Plants of both types are characterized by a graded control function between the vegetative and the reproductive phase. Most of the families showed a single switch from vegetative to reproductive growth, two families showed even two switch periods.Between-family differentiation is interpreted as a result of multiniche selection in populations of Erophila verna. Within-family variation is disadvantageous due to low fecundity.


Naringin Inhibits Colorectal Carcinogenesis by Inhibiting Viability of Colorectal Cancer Cells.
To explore the therapeutic effect of naringin on colorectal cancer (CRC) and the related mechanism. Cell counting kit-8 (CCK-8) assay and annexin V-FITC/PI assay were used to detect the effect of naringin (50-400 µg/mL) on cell proliferation and apoptosis of CRC cells, respectively. The scratch wound assay and transwell migration assay were used to assess the effect of naringin on CRC cell migration. Four-week-old male nude mice were injected with HCT116 cells subcutaneously to establish the tumor xenograft model. Naringin was injected intraperitoneally at 50 mg/(kg·d), with solvent and 5-fluorouracil treatment as control. The width and length of the tumors were measured and recorded every 6 days, and tumor tissues were photographed and weighed on the last day of the 24-d observation period. Immunohistochemical staining for caspase-3, proliferating cell nuclear antigen and TUNEL assay were used to evaluate the effect of naringin on cell proliferation and apoptosis in tumor tissues. The body weight, food and water intake of mice were recorded, and the major organs in different treatment groups were weighed on the last day and stained with hematoxylin and eosin for histological analysis. Meanwhile, the routine blood indicators were recorded. CCK-8 and annexin V-FITC/PI results confirmed that naringin (100, 200, and 400 µg/mL) could inhibit proliferation and promote apoptosis. The scratch wound assay and transwell migration assay results confirmed the inhibitory activity of naringin against CRC cells migration. In vivo results demonstrated the inhibitory effect of naringin on tumor growth with good bio-compatibility. Naringin inhibited colorectal carcinogenesis by inhibiting viability of CRC cells.


In vivo and in vitro formation of the junctional complex in choroid epithelium. A freeze-etching study.
The junctional complex of choroid epithelial cells was studied during in vivo formation, disaggregation after trypsin treatment, and in vitro reaggregation. The in vivo formation begins with the occurrence of amorphous patches of particles followed by the formation of small particulate rows and polygonal-ordered particle assemblies. Further arrangement of the zonula occludens continues with the confluence of particles and smooth contoured ridges. At the 9th day stage a fully developed zonula occludens has developed. In a subsequent step nexus become integrated within the tight junction formation. Disaggregation after trypsination results in fragmentation of the zonulae occludentes. Parts of the disassembling aggregates become incorporated in vacuoles indicating an endocytotic mode of "digestion". The in vitro reconstruction of the zonula occludens proceeds from remnants of the former zonula occludens. On the 3rd to 4th day of cultivation mature tight junctions are visible. In vitro integrations of nexus were observed during a later phase. On the 7th day, cultivated choroid epithelial cells reveal well differentiated junctional complexes consisting of continuous zonulae occludentes and integrated gap junctions.


Increased afferent vagal responses produced by epicardial application of nicotine on the canine posterior left ventricle.
Vagally mediated vasodepressor reflexes are observed in dogs and patients with acute myocardial ischemia or infarction and may be related to the Bezold-Jarisch reflex. To test whether nicotine and bradykinin produced site-specific vagal vasodepressor and sympathetic vasopressor responses, respectively, we compared blood pressure and heart rate responses to these drugs applied topically at six epicardial sites of the canine left ventricle: midanterior, apical anterior, midlateral, apical lateral, midposterior, and apical posterior left ventricle. In alpha-chloralose-anesthetized dogs with open chests a felt pad soaked in bradykinin (25 micrograms) or nicotine (50 micrograms) was applied to the test sites. Previous studies indicated that epicardial application of these drugs elicited a sympathetically mediated pressor response and a vagally mediated depressor response, respectively. Vasopressor response to bradykinin did not differ among six test sites in 10 dogs. However, vasodepressor response to nicotine was greater in the midposterior left ventricular epicardium than at other test sites in 16 dogs. A hypertensive response after the initial hypotensive response to nicotine was observed in the midanterior left ventricle and not in any other site. These data indicate that vagal vasodepressor responses to epicardial nicotine are greater in the canine midposterior left ventricle and are compatible with the higher incidence of bradycardia or hypotension during acute inferoposterior myocardial infarction in patients. Sympathetic vasopressor responses to epicardial bradykinin are not site specific. Half of the dogs showed no response to bradykinin or nicotine.


TDP-43 high throughput screening analyses in neurodegeneration: advantages and pitfalls.
Dysfunctions in RNA processing and in particular the aberrant regulation of RNA binding proteins (RBPs) have recently been shown to play a fundamental role in the pathogenesis of neurodegenerative diseases. Understanding the pathogenic mechanisms involved will require the elucidation of the role(s) played by these RBPs in the general cell metabolism and neuronal survival in particular. In the past, the preferred approach has been to determine first of all the functional properties of the factor(s) of interest and then use this knowledge to determine targets in biologically relevant events. More recently, novel experimental approaches such as microarrays, RNA-seq and CLIP-seq have also become very popular to study RBPs. The advantage of these approaches, collectively known as high throughput screening (HTS), is their ability to determine gene expression changes or RNA/protein targets at a global cellular level. In theory, HTS strategies should be ideal for uncovering novel functional roles/targets of any RBP inside the cell. In practice, however, there are still difficulties in getting a coherent picture from all the huge amount of data they generate, frequently not validated experimentally and thus of unknown value. They may even act unfavorably towards a specific increase of knowledge of RBP functions, as the incomplete results are taken as solid data. In this work we will illustrate as an example the use of the HTS methodologies to characterize the interactions of a specific RBP: TDP-43. The multiple functions of this protein in RNA processing and its involvement in the pathogenesis of several forms of amyotrophic lateral sclerosis, frontotemporal lobar degeneration and other neurodegenerative diseases make it an excellent substrate for our analysis of the various advantages and limitations of different HTS experimental approaches.


Secondary metabolites with new medicinal functions from marine organisms.
We have focused on the identification of natural key compounds that possess biologically and medicinally intriguing functions. Some of bioactive naturally occurring compounds isolated from marine organisms were found to possess unique biological activities. Halichlorine from the marine sponge Halichondria okadai was shown to inhibit the activity of nuclear factor-κB in endothelial cells and block L-type Ca²⁺ channels. Thus, it may have therapeutic potentials for diseases such as atherosclerosis and hypertension.


Endothelin receptors in the brain modulate autonomic responses and arrhythmogenesis during acute myocardial infarction in rats.
Endothelin has been implicated in various processes in the brain, including the modulation of sympathetic responses. The present study examined the pathophysiologic role of brain endothelin-receptors in the setting of acute myocardial infarction, characterized by high incidence of ventricular tachyarrhythmias. We investigated the effects of intracerebroventricular administration of antagonists of endothelin-receptors ETA, ETB, or both, during a 24 h-observation period post-coronary ligation in (n = 70) rats. Continuous recording was performed via implanted telemetry transmitters, followed by arrhythmia-analysis and calculation of autonomic indices derived from heart rate variability. The regional myocardial electrophysiologic properties were assessed by monophasic action potentials and multi-electrode recordings. Sympathetic-activity was decreased and vagal-activity was enhanced after intracerebroventricular ETA-receptor blockade, thus attenuating regional myocardial repolarization inhomogeneity. As a result, the incidence of ventricular tachyarrhythmias was markedly lower in this group. Such effects were also observed after intracerebroventricular blockade of ETB-, or both, ETA- and ETB-receptors, although to a lesser extent. ETA-receptors in the brain modulate sympathetic and vagal responses and alter arrhythmogenesis during evolving myocardial necrosis in rats. These findings provide insights into arrhythmogenic mechanisms during acute myocardial infarction and call for further investigation on the role of endothelin in the central autonomic network.


Effects of public policies in the prevention of cardiovascular diseases: a systematic review of global literature.
Given the growing interest worldwide in applying public policies to improve human health, we undertook a systematic review of studies investigating whether public policies targeting unhealthy products could reduce cardiovascular diseases. This study was a systematic review of the literature. We searched research studies published in 2000-2020 from major databases, including MEDLINE and Embase. We followed Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines and narratively synthesized the studies based on vote counting and direction of the intervention effect. Ninety-eight studies, mostly from high-income countries, met the inclusion criteria. Most studies were on public policies targeting sugar-sweetened beverages and tobacco, followed by alcohol, sugar, salt, and junk foods. Overall, many reported that several fiscal, regulatory, and educational policies generated beneficial effects of reducing the diseases. Those studies that reported no or limited effects highlighted several sociodemographic and health risk characteristics and design and implementation aspects of the policy interventions as factors limiting the policy effects; most of these are modifiable with appropriate policy interventions. For instance, low magnitude of tax, substitution with other unhealthy products, firms' competitive response strategies, pre-existence of smoking bans, incremental enactment of smoking regulations, degree of enforcement, and various sociocultural factors minimized the effects of the policies. The literature supports a growing consensus on the beneficial effects of public policy for improving human health. The design and implementation of public policies must address various impeding factors and incorporate appropriate remedial measures. Further research is needed from low- and middle-income countries and on whether and how multiple policy instruments work in tandem.


'The real stuff': implications for nursing of assessing and measuring a terminally ill person's quality of life.
Two quality of life (QoL) assessment and measurement tools, the Client Generated Index (CGI) and the McGill Quality of Life (MQOL) questionnaires, were trialled within district nursing palliative care to test usefulness and feasibility for holistic intervention selection, individualized palliative care planning, and measurement of the quality of dying. The specific focus of this paper is to discuss the less tangible outcomes of the trial, which illuminate the partly 'hidden' value and nature of clinical nursing. These outcomes include awareness that the use of such tools may: by actual administration of the tool be, in and of itself, a therapeutic nursing action; focus on 'the real stuff from the client's perspective, that which matters most to the terminally ill client, but may not be classically considered as prompting nursing intervention; and facilitate 'the real stuff' of nursing, perhaps known but not usually articulated by nurses, and which usually does not feature on care plans nor in time allocation schedules.


Genetic aberrations of the K-ras proto-oncogene in bladder cancer in Kashmiri population.
To assess the frequency of specific point mutations in the K-ras gene in a group of Kashmiri patients with bladder cancer. We analyzed the incidence of K-ras exon 1 gene mutations in tumors and surgical margins in 60 patients with transitional cell carcinoma of varied clinical stages and histological grades using the polymerase chain reaction-single strand conformation polymorphism and DNA sequencing. A significant correlation was found between the K-ras, the lymph node status, and tumor recurrence (P < 0.05). Also, smokers and patients with higher tumor grade showed a significantly higher relative risk of developing K-ras mutations than the normal ones. K-ras exon 1 gene mutations were found with low frequency in the bladder cancer tumors from Kashmir valley, which suggests that K-ras gene might be involved in a sub-set of bladder tumors, but it needs further investigation on a larger cohort sample to authenticate the current findings.


Occurrence of hepatitis A and E and norovirus GI and GII in ready-to-eat vegetables in Italy.
Fresh vegetables and their ready-to-eat (RTE) salads have become increasingly recognized as potential vehicles for foodborne diseases. The EU Reg. 1441/2007 establishes microbiological criteria for bacterial pathogens for products placed on the market during their shelf-life (i.e. Salmonella spp., Listeria monocytogenes) for pre-cut fruits and vegetables (RTE) whilst it does not address the problem of contamination by enteric viruses. In this study we investigated the contamination by hepatitis A virus (HAV), hepatitis E virus (HEV) and norovirus (NoV) in 911 ready-to-eat vegetable samples taken from products at retail in Apulia and in Lombardia. The vegetable samples were tested using validated real-time PCR (RT-qPCR) assays, ISO standardized virological methods and ISO culturing methods for bacteriological analysis. The total prevalence of HAV and HEV was 1.9% (18/911) and 0.6% (6/911), respectively. None of the samples analysed in this study was positive for NoV, Salmonella spp. or Listeria monocytogenes. The detection of HAV and HEV in RTE salads highlights a risk to consumers and the need to improve production hygiene. Appropriate implementation of hygiene procedures is required at all the steps of the RTE vegetable production chain and this should include monitoring of emerging viral pathogens.


Heavy metal monitoring, analysis and prediction in lakes and rivers: state of the art.
Several research efforts have been conducted to monitor and analyze the impact of environmental factors on the heavy metal concentrations and physicochemical properties of water bodies (lakes and rivers) in different countries worldwide. This article provides a general overview of the previous works that have been completed in monitoring and analyzing heavy metals. The intention of this review is to introduce the historical studies to distinguish and understand the previous challenges faced by researchers in analyzing heavy metal accumulation. In addition, this review introduces a survey on the importance of time increment sampling (monthly and/or seasonally) to comprehend and determine the rate of change of different parameters on a monthly and seasonal basis. Furthermore, suggestions are made for future research to achieve more understandable figures on heavy metal accumulation by considering climate conditions. Thus, the intent of the current study is the provision of reliable models for predicting future heavy metal accumulation in water bodies in different climates and pollution conditions so that water management can be achieved using intelligent proactive strategies and artificial neural network (ANN) techniques.


Kinetic analysis of UV-induced incision discriminates between fibroblasts from different xeroderma pigmentosum complementation groups, XPA heterozygotes and normal individuals.
The capacity of a variety of human fibroblasts to incise DNA following exposure to far ultraviolet-light is determined from the rate of single-strand DNA break accumulation in the presence of DNA synthesis inhibitors. We have quantitated incision, one of the early steps in the UV excision repair pathway, in cells form normal, xeroderma pigmentosum groups C, D, G, H and variant individuals, and in the parents of one XPA patient. On the basis of the estimated initial rates of incision the different XP cells examined in this work can be ranked as follows: XP variant much greater than XPH greater than XPH greater than XPD greater than XPC greater than XPG greater than XPA. In each cell strain breaks accumulate immediately after irradiation over a range of 0.5-20 Jm-2 with the exception of the XPC strain examined, where there is an initial delay of 15 min. The rate of incision in XPA heterozygote cells is roughly half that of normal fibroblasts. Analysis of the kinetics of break accumulation over short intervals after irradiation permits estimation of the apparent enzymatic parameters, Km and Vmax, for the incision step. The approximate values of Km and Vmax for normal and XP variant are similar while for the heterozygotes of an XPA individual Km values are normal (around 1 Jm-2), but there is only half the amount of normal enzyme activity. XPD and H cells express low levels of active enzyme, between 5 and 15% of that of the normal, but while the Km of XPH is very similar to that of normal cells, that of two XPD strains examined is between 2- and 3-fold higher.


Differential scanning calorimetry as a complementary diagnostic tool for the evaluation of biological samples.
Differential scanning calorimetry (DSC) is a tool for measuring the thermal stability profiles of complex molecular interactions in biological fluids. DSC profiles (thermograms) of biofluids provide specific signatures which are being utilized as a new diagnostic approach for characterizing disease but the development of these approaches is still in its infancy. This article evaluates several approaches for the analysis of thermograms which could increase the utility of DSC for clinical application. Thermograms were analyzed using localized thermogram features and principal components (PCs). The performance of these methods was evaluated alongside six models for the classification of a data set comprised of 300 systemic lupus erythematosus (SLE) patients and 300 control subjects obtained from the Lupus Family Registry and Repository (LFRR). Classification performance was substantially higher using the penalized algorithms relative to localized features/PCs alone. The models were grouped into two sets, the first having smoother solution vectors but lower classification accuracies than the second with seemingly noisier solution vectors. Coupling thermogram technology with modern classification algorithms provides a powerful diagnostic approach for analysis of biological samples. The solution vectors from the models may reflect important information from the thermogram profiles for discriminating between clinical groups. DSC thermograms show sensitivity to changes in the bulk plasma proteome that correlate with clinical status. To move this technology towards clinical application the development of new approaches is needed to extract discriminatory parameters from DSC profiles for the comparison and diagnostic classification of patients.


Regulation of Stem Cell Self-Renewal and Oncogenesis by RNA-Binding Proteins.
Throughout their life span, multicellular organisms rely on stem cell systems. During development pluripotent embryonic stem cells give rise to all cell types that make up the organism. After birth, tissue stem cells maintain properly functioning tissues and organs under homeostasis as well as promote regeneration after tissue damage or injury. Stem cells are capable of self-renewal, which is the ability to divide indefinitely while retaining the potential of differentiation into multiple cell types. The ability to self-renew, however, is a double-edged sword; the molecular mechanisms of self-renewal can be a target of malignant transformation driving tumor development and progression. Growing lines of evidence have shown that RNA-binding proteins (RBPs) play pivotal roles in the regulation of self-renewal by modulating metabolism of coding and non-coding RNAs both in normal tissues and in cancers. In this review, we discuss our current understanding of tissue stem cell systems and how RBPs regulate stem cell fates as well as how the regulatory functions of RBPs contribute to oncogenesis.


Effect of D3 dopamine receptor on dopamine D4 receptor expression and function in renal proximal tubule cells from Wistar-Kyoto rats and spontaneously hypertensive rats.
Dopamine receptors induce natriuresis in kidney. Previous studies have shown interactions between different subtypes of dopamine receptors in renal proximal tubule (RPT) cells. We hypothesize that D3 receptors have an interaction with D4 receptors in RPT cells from normotensive rats (Wistar-Kyoto, WKY) and spontaneously hypertensive rats (SHRs). Immunoblotting and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to examine the expression of D3 and D4 receptors. Na-K-ATPase activity was used to measure the function of receptors. The distribution and colocalization of D3 and D4 receptors were detected by confocal microscopy and co-immunoprecipitation. D3 receptor agonist PD128907 increased the mRNA and protein expression of D4 receptors in RPT cells from WKY rats, but decreased that from SHRs. In the presence of PLC blocker (U73122, 10-mol/l) or PKC inhibitor 19 -31 (10-mol/l), the up-regulation of D3 receptor on D4 receptor was lost in WKY cells. Moreover, stimulation with PD128907 for 30 minutes decreased D4 receptor degradation in WKY cells, not in SHR cells. D3 and D4 receptors colocalized and co-immunoprecipitated in RPT cells. PD128907 increased co-immunoprecipitation of D3 and D4 receptors in WKY RPT cells, but not in SHR RPT cells. Pre-treatment with D3 receptor agonist also increases D4 receptor mediated inhibitory effect on Na-K-ATPase activity in WKY cells, but not in SHR cells. Renal D3 receptor regulates the expression and function of D4 receptor in RPT cells via PLC /PKC signaling pathway, the loss of this interaction might be involved in the pathogenesis of hypertension.


Osteopontin and bone sialoprotein distribution at the bone graft recipient site.
The area in close proximity to a bone graft is subject to marked remodeling activity, which may dramatically affect graft survival. To specifically analyze the effects at the recipient bed-onlay graft interface. In 22 adult Lewis rats, bicortical grafts were positioned below the temporal muscle and subperiosteally over the parietal bone. The recipient bone was left intact or ground to remove the external cortical layer, thereby exposing the graft to the osteopotent cells of the bone marrow. The rats were killed after 4 or 20 weeks. The outcome was assessed by routine histologic examination and immunohistochemical labeling for 2 bone matrix proteins, osteopontin and bone sialoprotein, which are involved in bone resorption and formation, respectively. Placement of the grafts submuscularly or grinding of the outer cortical layer of the host bed increased recipient site resorption. Resorptive activity (labeling) was concentrated to a subzone below the surface of the recipient bone; neither the graft surface nor the interface soft tissues were labeled. The successive loss of skeletal contour after bone grafting, in many cases, may largely result from recipient site failure rather than graft size reduction.


Dietary polyphenols identified as intracellular protein kinase A inhibitors.
Dietary plants contain several thousands different polyphenols that can potentially influence normal and pathological cellular processes through modulation of intracellular signaling pathways. A few polyphenols have been shown to be potent inhibitors of protein kinases. To identify possible dietary protein kinase A (PKA) inhibitors we designed a method for screening of substances in crude mixtures of food items for modulation of intracellular PKA activity that enables high-throughput testing of a large number of compounds and extracts. Luciferase was mutated to render it sensitive to phosphorylation by PKA (luciferase(PKA)) and transfected into a human hepatoma cell line (HepG2). Cells were then treated with extracts from dietary plants, including berries, fruits and spices, and intracellular PKA-activity was assessed by change in bioluminescence in live cells by imaging. Several extracts were found to inhibit PKA activity in a 96-well platform high-throughput screen. Green tea, crowberry, clove and cinnamon extracts were found to reduce intracellular cAMP levels consistent with their ability to increase luminescence from luciferase(PKA). Also pomegranate extract inhibited intracellular PKA and was used to estimate cellular association of polyphenols by HPLC and LC-MS. Pomegranate extract contains several anthocyanins, including delphinidin-3 glucoside. Delphinidin aglycone was found to inhibit cellular PKA activity in a concentration dependent manner. The inhibitory activity was found to be structure specific as a closely related compound to delphinidin had no activity. The current work identify phytochemicals in crude extracts which modulate cell signaling through PKA in a way that facilitate high through-put screening to help elucidate how plant based diet reduce risks of chronic diseases.


Novel mutations of CETP gene in Italian subjects with hyperalphalipoproteinemia.
Cholesteryl ester transfer protein (CETP) is a plasma glycoprotein that catalyses the transfer of cholesteryl esters from HDL to the other plasma lipoproteins. Genetic deficiency of CETP is one of the known causes of elevation of plasma HDL-C (primary hyperalphalipoproteinemia, HALP). We sequenced CETP gene in a group of 24 Italian subjects with primary HALP (HDL-C>80 mg/dl) suspected to have CETP deficiency. Two unrelated subjects both coming from the same geographical district, were found to be heterozygous for a nucleotide substitution in exon 6 (c.544C>T) and another subject was found to be heterozygous for a C>T transition in exon 9 (c.802C>T). Both mutations introduce a premature stop codon and are predicted to cause the production of truncated proteins (Q165X and R268X, respectively) devoid of function. The fourth proband was found to carry a T>C substitution in intron 15 (c.1407+2T>C) predicted to abolish the function of the donor splice site. To define the effect of this mutation on CETP pre-mRNA splicing we analysed CETP mRNA in COS-1 cells expressing a CETP minigene harbouring the mutation. The analysis of minigene transcript in COS-1 cells showed that IVS15+2T>C mutation caused the formation of an abnormal mRNA in which exon 14 joins directly to exon 16, predicted to encode a truncated peptide of 435 amino acids. In mutation carriers plasma CETP activity was found to be reduced by 38-60%. These are the first mutations in the CETP gene found in Italian subjects with HALP.


Studying isoform-specific inositol 1,4,5-trisphosphate receptor function and regulation.
Inositol 1,4,5-trisphosphate receptors (InsP3R) are a family of ubiquitously expressed intracellular Ca2+ channels. Isoform-specific properties of the three family members may play a prominent role in defining the rich diversity of the spatial and temporal characteristics of intracellular Ca2+ signals. Studying the properties of the particular family members is complicated because individual receptor isoforms are typically never expressed in isolation. In this article, we discuss strategies for studying Ca2+ release through individual InsP3R family members with particular reference to methods applicable following expression of recombinant InsP3R and mutant constructs in the DT40-3KO cell line, an unambiguously null InsP3R expression system.


Mechanistic consideration of the effect of perioperative volatile anesthetics on phagocytes.
A growing literature has shown that volatile anesthetics are promiscuous molecules targeting multiple molecules, some of which are critical for immunological functions. We focused on studies that delineated target molecules of volatile anesthetics on immune cells and summarized the effects of volatile anesthetics on immune functions. We also presented the perspectives of studying volatile anesthetics-mediated immunomodulation.


Markers of platelet apoptosis: methodology and applications.
Apoptosis, or programmed cell death, is a physiological mechanism that serves for controlled deletion of damaged cells. While long attributed exclusively to nucleated cells, over recent years it has been recognized that apoptosis also occurs in anucleate platelets. We describe here experiences of determining markers of apoptosis in human platelets treated in vitro with pro-apoptotic chemical and physical stimuli. These include depolarization of mitochondrial inner membrane, cytochrome c release, expression of pro-apoptotic and anti-apoptotic proteins of Bcl-2 family, activation of apoptosis executioner caspase-3, exposure of phosphatidylserine, platelet shrinkage, fragmentation to microparticles, blebbing and filopod extrusion on the platelet surface. These assays serve to characterize platelet apoptosis in different cellular compartments (mitochondria, cytosol and plasma membrane) and at the whole-cell level. Methods commonly employed in studies of platelet apoptosis markers include flow cytometry, Western blot analysis and electron microscopy. An integrated methodological approach, based on determination of different platelet apoptosis markers, represents a useful tool for examining platelet apoptosis in various physiological and pathological settings.


Trypan blue- and indocyanine green-assisted epiretinal membrane surgery: clinical and histopathological studies.
To evaluate the clinical outcome and electron microscopic findings of trypan blue (Tb) and indocyanine green (ICG) assisted epiretinal membrane (ERM) surgery. This is a prospective consecutive noncomparative interventional case series. After pars plana vitrectomy, 0.1 ml of 0.6 mg/ml Tb solution was applied for 1 min under air for ERM staining. After ERM removal, internal limiting membrane (ILM) was further peeled after staining with 0.2 ml of 1 mg/ml ICG solution. Intraoperative specimens were sent for electron microscopy. Tb was considered useful if the edge of ERM was stained where peeling could be initiated with a clearer visualisation of the overall extent of the ERM. In all, 16 eyes from 16 patients were recruited. There were nine grade 1 ERMs, five grade 2 ERMs, and two grade 3 ERMs. Tb was useful in six (67%) of the nine eyes with grade 1 ERMs and in all eyes with grade 2 or 3 ERMs. The three remaining grade 1 ERMs were removed together with surrounding ILM that was stained by ICG. The mean line of improvement was 1.3 lines with the median BCVA improved from 6/12 to 6/9. All 16 eyes had symptomatic improvement and none developed ERM recurrence. No complication related to Tb or ICG was observed clinically or angiographically. Electron microscopy of the Tb-stained ERM specimens showed fragments of ILM in all specimens. Tb and ICG are useful intraoperatively to improve the visualisation and facilitate complete removal of ERM and ILM in macular ERM surgery.


Benign EEG for prognostication of favorable outcome after cardiac arrest: A reappraisal.
The current EEG role for prognostication after cardiac arrest (CA) essentially aims at reliably identifying patients with poor prognosis ("highly malignant" patterns, defined by Westhall et al. in 2014). Conversely, "benign EEGs", defined by the absence of elements of "highly malignant" and "malignant" categories, has limited sensitivity in detecting good prognosis. We postulate that a less stringent "benign EEG" definition would improve sensitivity to detect patients with favorable outcomes. Retrospectively assessing our registry of unconscious adults after CA (1.2018-8.2021), we scored EEGs within 72 h after CA using a modified "benign EEG" classification (allowing discontinuity, low-voltage, or reversed anterio-posterior amplitude development), versus Westhall's "benign EEG" classification (not allowing the former items). We compared predictive performances towards good outcome (Cerebral Performance Category 1-2 at 3 months), using 2x2 tables (and binomial 95% confidence intervals) and proportions comparisons. Among 381 patients (mean age 61.9 ± 15.4 years, 104 (27.2%) females, 240 (62.9%) having cardiac origin), the modified "benign EEG" definition identified a higher number of patients with potential good outcome (252, 66%, vs 163, 43%). Sensitivity of the modified EEG definition was 0.97 (95% CI: 0.92-0.97) vs 0.71 (95% CI: 0.62-0.78) (p < 0.001). Positive predictive values (PPV) were 0.53 (95% CI: 0.46-0.59) versus 0.59 (95% CI: 0.51-0.67; p = 0.17). Similar statistics were observed at definite recording times, and for survivors. The modified "benign EEG" classification demonstrated a markedly higher sensitivity towards favorable outcome, with minor impact on PPV. Adaptation of "benign EEG" criteria may improve efficient identification of patients who may reach a good outcome.


Histopathological study on N-ethyl-N'-nitro-N-nitrosoguanidine-induced colon cancer in dogs.
We prepared a suppository containing 50 mg of N-ethyl-N'-nitro-N-nitrosoguanidine (ENNG), and successfully produced experimental colon cancer with good reproducibility by continuous intrarectal insertion of one or two suppositories per day in dogs. The tumors were very similar to human colon cancers, macroscopically and histologically. In one of three dogs subjected to histopathological study, metastases to the lymph nodes, lung, liver and kidney were observed. This animal model produced by a simple procedure will be helpful in investigating treatment of rectal cancer.


A natural biomineral for enhancing the biomineralization and cell response of 3D printed polylactic acid bone scaffolds.
Polylactic acid (PLA) has been extensively used as a bone scaffold material, but it still faces many problems including low biomineralization ability, weak cell response, low mechanical properties, etc. In this study, we proposed to utilize the distinctive physical, chemical and biological properties of a natural biomineral with organic matrix, pearl powder, to enhance the overall performance of PLA bone scaffolds. Porous PLA/pearl composite bone scaffolds were prepared using fused deposition modeling (FDM) 3D printing technology, and their comprehensive performance was investigated. Macro- and micro- morphological observation by the optical camera and scanning electron microscopy (SEM) showed the 3D printed scaffolds have interconnected and ordered periodic porous structures. Phase analysis by X-ray diffraction (XRD) indicated pearl powder was well composited with PLA without impurity formation during the melt extrusion process. The mechanical test results indicated the tensile and compressive strength of PLA/pearl composite scaffolds with 10 % pearl powder content yielded the highest values, which were 15.5 % and 21.8% greater than pure PLA, respectively. The water contact angle and water absorption tests indicated that PLA/pearl showed better hydrophilicity than PLA due to the presence of polar groups in the organic matrix of the pearl powder. The results of the simulated body fluid (SBF) soaking revealed that the addition of pearl powder effectively enhanced the formation and deposition of apatite, which was attributed to the release of Ca2+ from the dissolution of pearl powder. The cell culture of bone marrow mesenchymal stem cells (BMSCs) indicated that PLA/pearl scaffolds showed better cell proliferation and osteogenic differentiation than PLA due to the stimulation of the biological organic matrix in pearl powder. These outcomes signify the potential of pearl powder as a natural biomineral containing bio-signal factors to improve the mechanical and biological properties of polymers for better bone tissue engineering application.


Electrophysiological and electron-microscopical correlations with fluid and electrolyte secretion in rabbit ileum.
1. The effects of theophylline and triaminopyrimidine on the passive permeability of Na+ and Cl- across sheets of rabbit ileum treated with 0.1 mM-ouabain were examined by determining the NaCl: mannitol dilution potentials, K+:Na+; choline: Na+ and SO24-:Cl- biionic potentials. The results indicate (a) that triaminopyrimidine reduces paracellular Na+ and K+ permeability without affecting Cl- permeability and (b) that theophylline increases Cl- permeability without affecting Na+ permeability and (c) that neither theophylline nor triaminopyrimidine interfere with each other's action. This is further evidence consistent with separate routes for paracellular Na+ and Cl- movement. 2. Electrical resistance changes across sheets of actively transporting rabbit ileum were measured as a function of time in various conditions. Theophylline has a biphasic effect on resistance. Initially it decreases resistance from 56 omega cm2 (control) to 40 omega cm2. In the ensuing 30 min, resistance rises to 50 omega cm2; whereas it falls to 30 omega cm2 in controls. When theophylline is present, with triaminopyrimidine, or with galactose, no secondary rise in tissue resistance occurs. The initial decrease in resistance is consistent with a theophylline-dependent increase in Cl- conductance and the secondary rise in resistance may be attributed to collapse of the lateral intercellular space following leakage of NaCl and fluid into the mucosal solution. 3. Electron microscopy of glutaraldehyde-fixed tissue confirms the above views, since, with theophylline present, the lateral intercellular spaces are collapsed and with both triaminopyrimidine and theophylline, or both theophylline and 20 mM-galactose present the spaces remain open. 4. It is shown in the Discussion that the theophylline- or choleragen induced increase in passive Cl- permeability of the mucosal border is the only requirement necessary to explain the increase in electrogenic Cl- secretion, the increase in short-circuit current, as well as neutral secretion of NaCl and net fluid secretion.


A method for localizing atrial type B receptors in the dog.
1. The effect of ectopic stimulation of atria (premature ventricular contraction) on the activity of atrial type B receptors has been studied in dogs. 2. In sixteen open-chest dogs, discharge from right or left atrial type B receptors (identified by their response to pulmonary artery occlusion) was recorded. Direct stimulation of either atrium produced an increase in the activity of the left atrial but a decrease in the right atrial receptors. The earlier during the ventricular systole that the premature ventricular contraction occurred the more marked was the effect. 3. In a separate series of fourteen closed-chest experiments the right atrium was stimulated internally via the external jugular vein. The effects on the activity of the atrial type B receptors were similar to those observed during the open-chest experiments. 4. It is concluded that in the dog the technique of internal stimulation of the right atrium without opening the chest can be used to distinguish between the right and left atrial type B receptors.


Cell division inhibitors SulA and MinC/MinD block septum formation at different steps in the assembly of the Escherichia coli division machinery.
SulA and MinCD are specific inhibitors of cell division in Escherichia coli. In this paper, size exclusion chromatography was used to study the effect of the SulA and MinCD division inhibitors on the oligomerization state of endogenous FtsZ in cytoplasmic extracts, and immunofluorescence microscopy was used to determine the effect of SulA and MinCD on the formation of FtsZ, FtsA and ZipA rings at potential division sites. SulA prevented the formation of high-molecular-weight FtsZ polymers by interfering with FtsZ dimerization and subsequent oligomerization. In contrast, the MinCD division inhibitor did not prevent the oligomerization of FtsZ in the cell extracts or the formation of FtsZ and ZipA ring structures in vivo. However, MinCD did prevent the formation of FtsA rings. Increased expression of ftsA suppressed MinCD-induced division inhibition, but had no effect on SulA-induced division inhibition. These results indicate that MinCD blocks the assembly of the septation machinery at a later step than SulA, at the stage at which FtsA is added to the FtsZ ring.


Simultaneous determination of atorvastatin and valsartan in human plasma by solid-based disperser liquid-liquid microextraction followed by high-performance liquid chromatography-diode array detection.
A simple, sensitive, and efficient method has been developed for simultaneous estimation of valsartan and atorvastatin in human plasma by combination of solid-based dispersive liquid-liquid microextraction and high performance liquid chromatography-diode array detection. In the proposed method, 1,2-dibromoethane (extraction solvent) is added on a sugar cube (as a solid disperser) and it is introduced into plasma sample containing the analytes. After manual shaking and centrifugation, the resultant sedimented phase is subjected to back extraction into a small volume of sodium hydrogen carbonate solution using air-assisted liquid-liquid microextraction. Then the cloudy solution is centrifuged and the obtained aqueous phase is transferred into a microtube and analyzed by the separation system. Under the optimal conditions, extraction recoveries are obtained in the range of 81-90%. Calibration curves plotted in drug-free plasma sample are linear in the ranges of 5-5000μgL(-1) for valsartan and 10-5000μgL(-1) for atorvastatin with the coefficients of determination higher than 0.997. Limits of detection and quantification of the studied analytes in plasma sample are 0.30-2.6 and 1.0-8.2μgL(-1), respectively. Intra-day (n=6) and inter-days (n=4) precisions of the method are satisfactory with relative standard deviations less than 7.4% (at three levels of 10, 500, and 2000μgL(-1), each analyte). These data suggest that the method can be successfully applied to determine trace amounts of valsartan and atorvastatin in human plasma samples.


Predictive factors for response to anti-thymocyte globulin in acquired aplastic anemia.
Lymphocyte phenotype, bone marrow cellularity, in vitro marrow growth potential, and treatment responses were examined in 22 patients with acquired aplastic anemia who were given anti-thymocyte globulin. Eleven of 22 patients (50 percent) had a significant hematologic response within three months of therapy, confirming the effectiveness of this therapy. Pretreatment fetal hemoglobin concentration, age, sex, severity of pancytopenia, degree of maximal lymphopenia, and total hemolytic complement consumption did not affect response. Patients with severe disease who were treated within 16 weeks after diagnosis had a higher response rate (55 percent) than those treated after 16 weeks (20 percent). In patients with severe disease, a higher pretherapy bone marrow cellularity (p less than 0.001) and presence of granulocyte-macrophage colony growth correlated with response. The actuarial survival of patients with response in the group with severe disease was 100 percent at 12 months as compared with 33 percent in patients without response. Following anti-thymocyte globulin therapy, a reduced number of blood lymphocyte T subpopulations was seen in all patients. At three months after therapy, the patients with response who had severe disease had increased T8-positive cells (+56 percent versus -4 percent for patients without response) and la-positive cells (+32 percent versus -62 percent).


The effects of Na+ and K+ on the conformational transitions of (Na+ + K+)-ATPase.
The equilibrium between the E1 form and the E2 form of the (Na+ + K+)-ATPase is poised towards E2 in the absence of cations when monitored by the fluorescence of eosin. Na+ converts the enzyme to the E1 form with a K0.5 of 1.9 mM (pH 7.4 and 2 microM eosin). The titration curves indicate that more than one Na+ is necessary. K0.5 for K+ for reversal of the Na+ effect is 0.8 mM with 10 mM Na+, 5.8 mM K+ with 30 mM Na+ and 37.7 mM with 100 mM Na+. In the absence of K+ the rate of transition from E2 to E1 is rapid when Na+ is added, t1/2 is about 50 ms at 6 degrees C (pH 7.4). K+ in low concentrations decreases the rate with a K0.5 for K+ of about 10 microM, and t1/2 is about 2 s at 1 mM K+. K+ in concentrations from 1 to 50 mM (the highest tested) increases the rate of transition from E2 to E1. E2 thus consists of a non-K+ form (E2), a K+ both with K+ bound with apparent high affinity, the 'occluded' form (KoccE2), and a form with K+ bound both at high and low affinity sites (KE2). K+ bound to the low-affinity sites facilitates the transition from KoocE2 to E1. So does an increase in the Na+ concentration, but Na+ is less effective than K+. The rate of transition from E1 to E2 is rapid when K+ is added to enzyme in 20 mM Na+. The rate increases with the K+ concentration, and the effect of K+ does not saturate at the concentrations of K+ sufficient to displace the equilibrium fully to E2. Na+ decreases the rate of transition from E1 to E2. A model is suggested for the effect of the cations on the transition between the different forms of the enzyme. The physiological implications of the effect of K+ at a low-affinity site are discussed.


Identification of genetic loci controlling the characteristics and severity of brain and spinal cord lesions in experimental allergic encephalomyelitis.
Experimental allergic encephalomyelitis (EAE) is the principal genetically determined animal model for multiple sclerosis (MS), the major inflammatory disease of the central nervous system (CNS). Although genetics clearly play a role in susceptibility to MS, attempts to identify the underlying genes have been disappointing. Considerable variation exists between MS patients with regard to the severity of clinical signs, mechanism of demyelination, and location of CNS lesions, confounding the interpretation of genetic data. A mouse-human synteny mapping approach may allow the identification of candidate susceptibility loci for MS based on the location of EAE susceptibility loci. To date, 16 regions of the mouse genome have been identified that control susceptibility or clinical signs of EAE. In this work, we examined the genetic control of histopathological lesions of EAE in an F2 intercross population generated from the EAE susceptible SJL/J and EAE resistant B10.S/DvTe mouse strains. Composite interval mapping was used to identify 10 quantitative trait loci (QTL), including seven newly identified loci controlling the distribution and severity of CNS lesions associated with murine EAE. QTL on chromosome 10 control lesions in the brain, whereas QTL on chromosomes 3, 7, and 12 control lesions in the spinal cord. Furthermore, sexually dimorphic QTL on chromosomes 2, 9, and 11 control CNS lesions in females, whereas QTL on chromosomes 10, 11, 12, 16, and 19 control lesions in males. Our results suggest that the severity and location of CNS lesions in EAE are genetically controlled, and that the genetic component controlling the character and severity of the lesions can be influenced by sex.


Socioeconomic status and preterm birth: New Zealand trends, 1980 to 1999.
While a number of countries have reported rising preterm birth rates over the past two decades, none has examined the effects of socioeconomic status on preterm birth at a national level. To document the changing incidence of preterm birth in New Zealand over the past 20 years and to determine whether particular socioeconomic or ethnic subsections of the population have contributed disproportionately to the changes seen. Birth registration data routinely available from the New Zealand Health Information Service were analysed for the period 1980-99. Information for a total of 1 079 478 singleton live births was linked by Domicile Code to the New Zealand Deprivation Index, a small area index of deprivation. Singleton preterm birth rates rose by 37.2% during the 20 year period, from 4.3% in 1980 to 5.9% in 1999. Rates increased by 71.9% among those living in the most affluent areas, but by only 3.5% among those living in the most deprived areas, resulting in the disappearance of a socioeconomic gradient in preterm birth that had existed during the early 1980s. This study challenges traditional thinking on the associations between socioeconomic status and preterm birth. Further research is necessary if the changes that have occurred in New Zealand over the past 20 years are to be fully understood.


Perinatal and first year follow-up of patients with Prader-Willi syndrome: normal size of hands and feet.
Four patients with Prader-Willi syndrome, diagnosed in the neonatal period and followed during the first year of life, are reported. There were three males and one female. All four patients presented with hypotonia and distinct craniofacial dysmorphism. Prometaphase chromosome analysis showed interstitial deletion of 15q in all of them. The placentae and umbilical cords were examined in three of the patients and found normal. Electromyography done in the neonatal period suggested primary myopathy. Height, weight and head circumference were normal at birth in all patients. Hand and foot measurements showed normal size at birth and during the first year of life.


Parental attitudes in the pediatric emergency department about the COVID-19 vaccine.
COVID-19 vaccinations are now recommended in the United States (U.S.) for children ≥ 6 months old. However, pediatric vaccination rates remain low, particularly in the Hispanic/Latinx population. Using the 4C vaccine hesitancy framework (calculation, complacency, confidence, convenience), we examined parental attitudes in the emergency department (ED) towards COVID-19 vaccination, identified dimensions of parental vaccine hesitancy, and assessed parental willingness to have their child receive the COVID-19 vaccine. As part of a larger multi-methods study examining influenza vaccine hesitancy, we conducted interviews that included questions about COVID-19 vaccine authorization for children. We used directed content analysis to extract qualitative themes from 3 groups of parents in the ED: Hispanic/Latinx Spanish speaking (HS), Hispanic/Latinx English speaking (HE), non-Hispanic/non-Latinx White English speaking (WE). Themes were triangulated with the Parent Attitudes about Childhood Vaccines (PACV) survey, where higher scores indicate increased vaccine hesitancy. Factors influencing vaccine hesitancy were mapped to the 4C framework from 58 sets of interviews and PACVs. HE and HS parents, compared to WE parents, had less knowledge about COVID-19 and its vaccine, and more beliefs in COVID-19 vaccine myths. However, both HS and HE parent groups were more inclined to endorse COVID-19 vaccine effectiveness as a reason to have their children vaccinated. HS parents felt that COVID-19 increased their fear of illnesses in general and were worried about confusing COVID-19 with other infections. Median PACV scores of HS (Mdn = 20) and HE (Mdn = 20) parent groups were higher than of WE parents (Mdn = 10), but parental willingness to have their child receive COVID-19 vaccination was similar across groups. Higher COVID-19 vaccine hesitancy among HS and HE parents compared to WE parents may be attributed to insufficient knowledge about COVID-19, its vaccine, along with COVID-19 vaccine myths. Efforts to provide targeted vaccine education to different populations is warranted.


Diagnostic performance of whole-body [18F]FDG PET/MR in cancer M staging: A systematic review and meta-analysis.
To calculate the pooled diagnostic performances of whole-body [18F]FDG PET/MR in M staging of [18F]FDG-avid cancer entities. A diagnostic meta-analysis was conducted on the [18F]FDG PET/MR in M staging, including studies: (1) evaluated [18F]FDG PET/MR in detecting distant metastasis; (2) compared[ 18F]FDG PET/MR with histopathology, follow-up, or asynchronous multimodality imaging as the reference standard; (3) provided data for the whole-body evaluation; (4) provided adequate data to calculate the meta-analytic performances. Pooled performances were calculated with their confidence interval. In addition, forest plots, SROC curves, and likelihood ratio scatterplots were drawn. All analyses were performed using STATA 16. From 52 eligible studies, 2289 patients and 2072 metastases were entered in the meta-analysis. The whole-body pooled sensitivities were 0.95 (95%CI: 0.91-0.97) and 0.97 (95%CI: 0.91-0.99) at the patient and lesion levels, respectively. The pooled specificities were 0.99 (95%CI: 0.97-1.00) and 0.97 (95%CI: 0.90-0.99), respectively. Additionally, subgroup analyses were performed. The calculated pooled sensitivities for lung, gastrointestinal, breast, and gynecological cancers were 0.90, 0.93, 1.00, and 0.97, respectively. The pooled specificities were 1.00, 0.98, 0.97, and 1.00, respectively. Furthermore, the pooled sensitivities for non-small cell lung, colorectal, and cervical cancers were 0.92, 0.96, and 0.86, respectively. The pooled specificities were 1.00, 0.95, and 1.00, respectively. [18F]FDG PET/MR was a highly accurate modality in M staging in the reported [18F]FDG-avid malignancies. The results showed high sensitivity and specificity in each reviewed malignancy type. Thus, our findings may help clinicians and patients to be confident about the performance of [18F]FDG PET/MR in the clinic. Although [18F]FDG PET/MR is not a routine imaging technique in current guidelines, mostly due to its availability and logistic issues, our findings might add to the limited evidence regarding its performance, showing a sensitivity of 0.95 and specificity of 0.97. • The whole-body [18F]FDG PET/MR showed high accuracy in detecting distant metastases at both patient and lesion levels. • The pooled sensitivities were 95% and 97% and pooled specificities were 99% and 97% at patient and lesion levels, respectively. • The results suggested that 18F-FDG PET/MR was a strong modality in the exclusion and confirmation of distant metastases.


Recurrence Rate and Pattern of Perihilar Cholangiocarcinoma after Curative Intent Resection.
The aim of this study was to investigate the rate and pattern of recurrence after curative intent resection of perihilar cholangiocarcinoma (PHC). Patients were included from 2 prospectively maintained databases. Recurrences were categorized by site. Time to recurrence and recurrence-free survival (RFS) were estimated using the Kaplan-Meier method. Cox proportional hazards modeling was used to identify independent poor prognostic factors. Between 1991 and 2012, 306 consecutive patients met inclusion criteria. Median overall survival was 40 months. A recurrence was diagnosed in 177 patients (58%). An initial local recurrence was found in 26% of patients: liver hilum (11%), hepaticojejunostomy (8%), liver resection margin (8%), or distal bile duct remnant (2%). An initial distant recurrence was observed in 40% of patients: retroperitoneal lymph nodes (14%), intrahepatic away from the resection margin (13%), peritoneum (12%), and lungs (8%). Only 18% of patients had an isolated initial local recurrence. The estimated overall recurrence rate was 76% at 8 years. After a recurrence-free period of 5 years, 28% of patients developed a recurrence in the next 3 years. Median RFS was 26 months. Independent prognostic factors for RFS were resection margin, lymph node status, and tumor differentiation. Only node-positive PHC precluded RFS beyond 7 years. Perihilar cholangiocarcinoma will recur in most patients (76%) after resection, emphasizing the need for better adjuvant strategies. The high recurrence rate of up to 8 years justifies prolonged surveillance. Only patients with an isolated initial local recurrence (18%) may have benefited from a more extensive resection or liver transplantation. Node-positive PHC appears incurable.


mTOR-dependent TFEB activation and TFEB overexpression enhance autophagy-lysosome pathway and ameliorate Alzheimer's disease-like pathology in diabetic encephalopathy.
Diabetic encephalopathy (DE) is a complication of type 2 diabetes mellitus (T2DM) that features Alzheimer's disease (AD)-like pathology, which can be degraded by the autophagy-lysosome pathway (ALP). Since transcription factor EB (TFEB) is a master regulator of ALP, TFEB-mediated ALP activation might have a therapeutic effect on DE, but this has yet to be investigated. We established T2DM mouse models and cultured HT22 cells under high-glucose (HG) conditions to confirm the role of ALP in DE. To further investigate this, both mice and HT22 cells were treated with 3-methyladenine (3-MA). We also analyzed the content of TFEB in the nucleus and cytoplasm to evaluate its role in ALP. To confirm the effect of TFEB activation at the post-translational level in DE, we used rapamycin to inhibit the mechanistic target of rapamycin (mTOR). We transduced both mice and cells with TFEB vector to evaluate the therapeutic effect of TFEB overexpression on DE. Conversely, we conducted TFEB knockdown to verify its role in DE in another direction. We found that T2DM mice experienced compromised cognitive function, while HG-cultured HT22 cells exhibited increased cell apoptosis. Additionally, both T2DM mice and HG-cultured HT22 cells showed impaired ALP and heavier AD-like pathology. This pathology worsened after treatment with 3-MA. We also observed decreased TFEB nuclear translocation in both T2DM mice and HG-cultured HT22 cells. However, inhibiting mTOR with rapamycin or overexpressing TFEB increased TFEB nuclear translocation, enhancing the clearance of ALP-targeted AD-like pathology. This contributed to protection against neuronal apoptosis and alleviation of cognitive impairment. Conversely, TFEB knockdown lessened ALP-targeted AD-like pathology clearance and had a negative impact on DE. Our findings suggest that impaired ALP is responsible for the aggravation of AD-like pathology in T2DM. We propose that mTOR-dependent TFEB activation and TFEB overexpression are promising therapeutic strategies for DE, as they enhance the clearance of ALP-targeted AD-like pathology and alleviate neuronal apoptosis. Our study provides insight into the underlying mechanisms of DE and offers potential avenues for the development of new treatments for this debilitating complication of T2DM. Video abstract.


Surgeon distress as calibrated by hours worked and nights on call.
The relationships of working hours and nights on call per week with various parameters of distress among practicing surgeons have not been previously examined in detail. More than 7,900 members of the American College of Surgeons responded to an anonymous, cross-sectional survey. The survey included self-assessment of their practice setting, a validated depression screening tool, and standardized assessments of burnout and quality of life. There was a clear gradient between hours and burnout, with the prevalence of burnout ranging from 30% for surgeons working <60 hours/week, 44% for 60 to 80 hours/week, and 50% for those working >80 hours/week (p < 0.001). When correlated with number of nights on call, burnout exhibited a threshold effect at ≥2 nights on call/week (≤1 nights on call/week, 30%; ≥2 nights on call/week, 44% to 46%; p < 0.0001). Screening positive for depression rate also correlated strongly with hours and nights on call (both p < 0.0001). Those who worked >80 hours/week reported a higher rate of medical errors compared with those who worked <60 hours/week (10.7% versus 6.9%; p < 0.001), and were twice as likely to attribute the error to burnout (20.1% versus 8.9%; p = 0.001). Not surprisingly, work and home conflicts were higher among surgeons who worked longer hours or had ≥2 nights on call. A significantly higher proportion of surgeons who worked >80 hours/week or had >2 nights on call/week would not become a surgeon again (p < 0.0001). Number of hours worked and nights on call per week appear to have a substantial impact on surgeons, both professionally and personally. These factors are strongly related to burnout, depression, career satisfaction, and work and home conflicts.


Cognitive, emotional and neurochemical effects of repeated maternal separation in adolescent rats.
As an adverse early life experience, maternal separation (MS) induces profound neurochemical, cognitive and emotional dysfunction. Previous studies have reported that MS affected prepulse inhibition (PPI), anxiety-related behaviors, dopaminergic and serotonergic activity in adult rats, and in the present study, we investigated the effects of repeated (4h/day) maternal separation during postnatal days 1-21 on PPI and anxiety-related behaviors in an elevated plus maze, as well as dopamine D2 receptor (DRD2) and 5-HT1A receptor expression in the medial prefrontal cortex (mPFC), nucleus accumbens (NAc) and hippocampus in adolescent rats. Our findings show that repeated MS results in reduced PPI, increased anxiety-related behaviors, decreased DRD2 protein expression in the NAc and hippocampus, and decreased 5-HT1A protein expression in the mPFC and hippocampus in adolescent rats. These data further demonstrate that MS can be used as an animal model of neuropsychiatric disease.


Safety pharmacology investigations on the nervous system: An industry survey.
The Safety Pharmacology Society (SPS) conducted an industry survey in 2015 to identify industry practices as they relate to central, peripheral and autonomic nervous system ('CNS') drug safety testing. One hundred fifty-eight (158) participants from Asia (16%), Europe (20%) and North America (56%) responded to the survey. 52% of participants were from pharmaceutical companies (>1000 employees). Oncology (67%) and neurology/psychiatry (66%) were the most frequent target indications pursued by companies followed by inflammation (48%), cardiovascular (43%), metabolic (39%), infectious (37%), orphan (32%) and respiratory (29%) diseases. Seizures (67% of participants), gait abnormalities (67%), tremors (65%), emesis (56%), sedation (52%) and salivation (47%) were the most commonly encountered CNS issues in pre-clinical drug development while headache (65%), emesis/nausea (60%), fatigue (51%) and dizziness (49%) were the most frequent issues encountered in Phase I clinical trials. 54% of respondents reported that a standard battery of tests applied to screen drug candidates was the approach most commonly used to address non-clinical CNS safety testing. A minority (14% of all participants) reported using electroencephalography (EEG) screening prior to animal inclusion on toxicology studies. The most frequent group size was n=8 for functional observation battery (FOB), polysomnography and seizure liability studies. FOB evaluations were conducted in a dedicated room (78%) by blinded personnel (66%) with control for circadian cycle (55%) effects (e.g., dosing at a standardized time; balancing time of day across treatment groups). The rat was reported as the most common species used for seizure liability, nerve conduction and drug-abuse liability testing.


Efficient learning in children with rapid GABA boosting during and after training.
It is generally thought that children learn more efficiently than adults. One way to accomplish this is to have learning rapidly stabilized such that it is not interfered with by subsequent learning. Although γ-aminobutyric acid (GABA) plays an important role in stabilization, it has been reported that GABAergic inhibitory processing is not fully matured yet in children compared with adults. Does this finding indicate that more efficient learning in children is not due to more rapid stabilization? Here, we measured the concentration of GABA in early visual cortical areas in a time-resolved fashion before, during, and after visual perceptual learning (VPL) within subjects using functional MRS (fMRS) and then compared the concentrations between children (8 to 11 years old) and adults (18 to 35 years old). We found that children exhibited a rapid boost of GABA during visual training that persisted after training ended, whereas the concentration of GABA in adults remained unchanged. Moreover, behavioral experiments showed that children exhibited rapid development of resilience to retrograde interference, which indicates that children stabilize VPL much faster than adults. These results together suggest that inhibitory processing in children's brains is more dynamic and adapts more quickly to stabilize learning than in adults, making learning more efficient in children.


A pyroptosis-related gene signature predicts prognosis and immune microenvironment in hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is a highly malignant tumor with a very poor prognosis. Pyroptosis is an inflammatory form of cell death and plays an important role in cancer development. The prognostic value of pyroptosis-related genes (PRGs) in HCC has not been studied extensively. Unsupervised consensus clustering analysis was performed to identify two subtypes based on the expression profiles of prognostic PRGs in the The Cancer Genome Atlas (TCGA) database, and the differences between the two subtypes were compared. A prognostic model based on four PRGs was established by further least absolute shrinkage and selection operator (LASSO) Cox regression analysis and multivariate Cox regression analysis. Two subtypes (clusters 1 and 2) were identified by consensus clustering based on prognostic PRGs in HCC. Survival outcomes, biological function, genomic alterations, immune cell infiltration, and immune checkpoint genes were compared between the subtypes. Cluster 2 had a worse survival outcome than cluster 1. Cluster 2 was enriched for hallmarks of cancer progression, TP53 mutation, tumor-promoting immune cells, and immune checkpoint genes, which may contribute to the poor prognosis. A prognostic risk signature that predicted the overall survival (OS) of patients was constructed and validated. Consequently, a risk score was calculated for each patient. Combined with the clinical characteristics, the risk score was found to be an independent prognostic factor for survival of HCC patients. Further analysis revealed that the risk score was closely associated with the levels of immune cell infiltration and the expression profiles of immune checkpoint genes. Collectively, our study established a prognostic risk signature for HCC and revealed a significant correlation between pyroptosis and the HCC immune microenvironment.


Correlation among lung damage after radiation, amount of lipid peroxides, and antioxidant enzyme activities.
The correlation between lipid peroxidation and morphologic changes was examined in Sprague-Dawley rat lungs after 30 Gy single thoracic radiation. The rats were sacrificed every week until the end of the fifth week after radiation. The left lungs were used for the measurement of lipid peroxides and antioxidant enzymes activities. The right lungs were examined by light and electron microscopy. Amounts of lung lipid peroxides were within normal limits, and no cellular degenerative changes were observed in the lungs except for subendothelial and interstitial edema 2 weeks after radiation. Lipid peroxides drastically increased and marked degenerative cellular changes such as edematous swelling, vacuolation, and destruction of cell membranes occurred in the alveolar septa following the third week after radiation. The activities of catalase were significantly higher during the period from the second to the fifth week and those of superoxide dismutase and glutathione peroxidase increased at the end of the fifth week. Our results demonstrated that the acceleration of lipid peroxidation was well correlated with the morphologic expression of cell injury in the irradiated lungs.


Communication: Explicitly correlated formalism for second-order single-particle Green's function.
We present an explicitly correlated formalism for the second-order single-particle Green's function method (GF2-F12) that does not assume the popular diagonal approximation and describes the energy dependence of the explicitly correlated terms. For small and medium organic molecules, the basis set errors of ionization potentials of GF2-F12 are radically improved relative to GF2: the performance of GF2-F12/aug-cc-pVDZ is better than that of GF2/aug-cc-pVQZ, at a significantly lower cost.


Localization and fluctuations in quantum kicked rotors.
We address the issue of fluctuations, about an exponential line shape, in a pair of one-dimensional kicked quantum systems exhibiting dynamical localization. An exact renormalization scheme establishes the fractal character of the fluctuations and provides a method to compute the localization length in terms of the fluctuations. In the case of a linear rotor, the fluctuations are independent of the kicking parameter k and exhibit self-similarity for certain values of the quasienergy. For given k, the asymptotic localization length is a good characteristic of the localized line shapes for all quasienergies. This is in stark contrast to the quadratic rotor, where the fluctuations depend upon the strength of the kicking and exhibit local "resonances." These resonances result in strong deviations of the localization length from the asymptotic value. The consequences are particularly pronounced when considering the time evolution of a packet made up of several quasienergy states.


Correction to: Association Between Temporal Spatial Parameters and Overuse Injury History in Runners: A Systematic Review and Meta-analysis.
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Intrathecal therapy of leptomeningeal CEM T-cell lymphoma in nude rats with anti-CD7 ricin toxin A chain immunotoxin.
We have established a new xenogeneic animal model of leptomeningeal metastasis (LM) by intracisternal inoculation of human CEM T-cell lymphoma into nude rats, and used it to evaluate the anti-lymphoma efficacy of an anti-CD7 ricin A chain immunotoxin (DA7). In vitro incubation with 2 microg/ml DA7 for 72 h inhibited CEM cells by 90% in a trypan blue exclusion assay. To establish its anti-lymphoma activity, one and four days after cisternal inoculation of 10(6) CEM cells, eight animals each were treated cisternally with 10 microg DA7 in 50 microl PBS or sham-treated with 50 microl PBS. Histopathologically, all eight sham-treated and five of eight DA7 treated animals showed typical features of LM with multilayers of tumor cells along the whole subarachnoid space and the ventricular walls, as well as subependymal and diffuse parenchymal tumor cell infiltration. Three DA7 treated animals were free of tumor. Two of these animals were asymptomatic long-term survivors (> 90 days). The third tumor-free animal suddenly died on day 51. Histology revealed viral myocarditis. Median symptom-free survival was 51 days (range 29-90+ days) in DA7 treated and 34 days (range 29-87 days) in sham-treated animals (p = 0.12, log-rank test). Histologically, no signs of neurotoxicity or systemic toxicity was found. However, DA7 treated animals showed a tendency to a slower weight increase on days 6-28 after tumor cell inoculation. Our results indicate that this model is useful in studying leptomeningeal seeding and intracisternal treatment of lymphoma. The demonstrated anti-tumor effect of DA7 treatment deserves further evaluation especially regarding the application of DA7 in early stages of LM from T-cell lymphoma.


Food matters: changing dimensions of science and practice in the nutrition profession.
This qualitative research describes some of the social organization of the American nutrition profession set in place by an apparent competition between two kinds of knowledge-a science-based knowledge and another more experimental knowledge drawn from practice. The methodological approach is constructivist and uses the theoretical framework of sociologist Dorothy Smith. Multiple data sources include: interviews and bibliographic data from 23 purposefully selected subjects who taught or studied in a graduate-level Nutrition Program at Teacher College, Columbia University from 1937 to 1992; administrative reports of the Program; faculty writings; dissertations approved by the program; the author's twenty-five years of experience in the nutrition profession; and a series of diagrams drawn to interpret and describe the data. Subjects recounted their varied professional experience during the 55-year period as a part of a larger study. The research traces changes in the relationship between nutrition science and practice as shaped by underlying sciences (food, agriculture, nutrition and health), related industries (food and health care), and culturally-bound gender ideals. Data analysis was based on recommended strategies for an inductive, interpretive, ethnographic case study. Subjects identified three forms of the construct of food-as nutrients, as marketable products, and as nurturance. This research suggests that the nurturing properties of food, the knowledge this generates, and the practitioners associated with nurturance are vulnerable and risk disappearance inside the profession, due to a history of resistance to this aspect of our discipline. The author recommends collaborative research between nutritionists and historians, sociologists and others to bring forth a more critical history of this profession.


Linking primary and metastatic tumour re-initiation.
Whether the cancer cells responsible for the growth of primary tumours are also able to re-initiate tumour growth after seeding to distant organs is unclear. The characterization of breast cancer cells with both of these attributes now identifies the functional and molecular determinants necessary to mediate primary tumour formation and re-initiation at the secondary site.