On the physical origin of conical bubble structure under an ultrasonic horn.
The cavitation field generated by an ultrasonic horn at low frequency and high power is known to self-organize into a conical bubble structure. The physical mechanism at the origin of this bubble structure is investigated using numerical simulations and acoustic pressure measurements. The thin bubbly layer lying at horn surface is shown to act as a nonlinear thickness resonator that amplifies acoustic pressure and distorts acoustic waveform. This mechanism explains the self-stabilization of the conical bubble structure as well as the generation of shock wave and the focusing at very short distance.


Increased lead concentration in brain and potentiation of lead-induced neuronal depression in rats after combined treatment with lead and disulfiram.
The effects of disulfiram (tetraethylthiuram disulfide) on blood and brain lead levels and on lead-induced changes in growth and cerebellar Purkinje neuron excitability were assessed in adult Sprague-Dawley rats. Disulfiram is metabolized to diethyldithiocarbamate, which forms a lipophilic complex with lead, and can thereby influence the tissue distribution of lead. Pregnant rats were exposed to 0.25% lead acetate or an equimolar amount of sodium acetate in the drinking water, and these treatments were continued for 4 weeks after birth. Half of the mothers from each group were given 0.1 mmole/kg disulfiram orally twice a week until parturition, after which the treatment was continued for 4 weeks in the respective pups in the form of subcutaneous injections. Although lead exposure markedly increased blood lead levels, the increase in brain lead levels was much more modest. Disulfiram markedly increased brain lead levels while blood lead levels in this group were only slightly elevated as compared to animals receiving lead alone. In addition, the lead + disulfiram group had depressed weight gain during maturation, and Purkinje neuron firing rates were reduced. The lead alone and disulfiram alone groups were not different from controls in these respects. These data suggest that disulfiram potentiates the adverse effects of lead on growth rates and on cerebellar Purkinje neuron function by facilitating the accumulation of lead in brain tissue.


Music perception and speech intelligibility in noise performance by Italian-speaking cochlear implant users.
The goal of this study was to investigate the performance correlations between music perception and speech intelligibility in noise by Italian-speaking cochlear implant (CI) users. Twenty postlingually deafened adults with unilateral CIs (mean age 65 years, range 46-92 years) were tested with a music quality questionnaire using three passages of music from Classical Music, Jazz, and Soul. Speech recognition in noise was assessed using two newly developed adaptive tests in Italian: The Sentence Test with Adaptive Randomized Roving levels (STARR) and Matrix tests. Median quality ratings for Classical, Jazz and Soul music were 63%, 58% and 58%, respectively. Median SRTs for the STARR and Matrix tests were 14.3 dB and 7.6 dB, respectively. STARR performance was significantly correlated with Classical music ratings (rs = - 0.49, p = 0.029), whereas Matrix performance was significantly correlated with both Classical (rs = - 0.48, p = 0.031) and Jazz music ratings (rs = - 0.56, p = 0.011). Speech with competitive noise and music are naturally present in everyday listening environments. Recent speech perception tests based on an adaptive paradigm and sentence materials in relation with music quality measures might be representative of everyday performance in CI users. The present data contribute to cross-language studies and suggest that improving music perception in CI users may yield everyday benefit in speech perception in noise and may hence enhance the quality of listening for CI users.


Prognostic impact of a large mesenteric mass >2 cm in ileal neuroendocrine tumors.
Ileal neuroendocrine tumors (i-NETs) frequently metastasize to mesenteric lymph nodes and the liver. Regional lymphadenopathy is associated with desmoplasia of the mesentery forming a large mesenteric mass (LMM). Although the latest American Joint Committee on Cancer TNM staging (8th edition) defined LMM >2 cm as N2, the prognostic impact of LMM is ill-defined. We evaluated whether LMM is prognostic for patients with i-NETs. This single-institution, retrospective cohort study included 106 patients who underwent resection of i-NETs between 2007 and 2018. Overall survival (OS) and liver progression-free survival (LPFS) were compared between patients with and without LMM. LMM was present in 66 patients (62%) and was not associated with the presence or absence of liver metastasis (P = .969) or the extent of liver involvement (P = .938). OS and LPFS differed significantly between patients with and without LMM (5-year OS rates of 64.8% and 92.9%, respectively, P = .011; 3-year LPFS rates of 45.3% and 67.5%, respectively, P = .025). In multivariate analysis, LMM was an independent prognostic factor for both OS (hazard ratio: 4.69, 95% confidence interval: 1.63-17.6) and LPFS (1.99, 1.08-3.88). LMM >2 cm is prognostic for OS and LPFS and represents aggressive tumor biology.


Two-Year observational study of autonomic skin function in patients with Parkinson's disease compared to healthy individuals.
We characterized autonomic pilomotor and sudomotor skin function in early Parkinson's disease (PD) longitudinally. We enrolled PD patients (Hoehn and Yahr 1-2) and healthy controls from movement disorder centers in Germany, Hungary, and the United States. We evaluated axon-reflex responses in adrenergic sympathetic pilomotor nerves and in cholinergic sudomotor nerves and assessed sympathetic skin response (SSR), predominantly parasympathetic neurocardiac function via heart rate variability, and disease-related symptoms at baseline, after 2 weeks, and after 1 and 2 years. gov: NCT03043768. We included 38 participants: 26 PD (60% females, aged 62.4 ± 7.4 years, mean ± SD) and 12 controls (75% females, aged 59.5 ± 5.8 years). Pilomotor function was reduced in PD compared to controls at baseline when quantified via spatial axon-reflex spread (78 [43-143], median [interquartile range] mm2 vs. 175 [68-200] mm2 , p = 0.01) or erect hair follicle count in the axon-reflex region (8 [6-10] vs. 11 [6-16], p = 0.008) and showed reliability absent any changes from baseline to Week 2 (p = not significant [ns]). Between-group differences increased over the course of 2 years (p < 0.05), although no decline was observed within groups (p = ns). Pilomotor impairment in PD correlated with motor symptoms (rho = -0.59, p = 0.017) and was not lateralized (p = ns). Sudomotor axon-reflex and neurocardiac function did not differ between groups (p = ns), but SSR was reduced in PD (p = 0.0001). Impairment of adrenergic sympathetic pilomotor function and SSR in evolving PD is not paralleled by changes to cholinergic sudomotor function and parasympathetic neurocardiac function, suggesting a sympathetic pathophysiology. A pilomotor axon-reflex test might be useful to monitor PD-related pathology.


Effectiveness of hardening films on die stone.
1. Cyanoacrylate resins and resinous die hardeners effectively improved the hardness and abrasion resistance of stone dies. 2. Application of liquid resins followed by blotting and/or blowing with compressed air produced an improved surface without clinically significant dimensional change. The extent of film buildup was qualitatively related to the viscosity of the resin. 3. Application of multiple coats of resin or simply soaking the dies in resin without further blotting or other mode of removal of the excess produced a thick film which can obliterate surface detail.


Gradient-index lens rod based probe for office-based optical coherence tomography of the human larynx.
Optical coherence tomography (OCT) is an evolving noninvasive imaging modality that has been used to image the human larynx during surgical endoscopy. The design of a long gradient-index lens-based probe capable of capturing images of the human larynx by use of spectral domain OCT during a typical office-based laryngoscopy examination is presented. An optical-ballast-based 4f optical relay system is proposed to realize variable working distance with a constant optical delay. In-vivo OCT imaging of the human larynx is demonstrated. Office-based OCT is a promising imaging modality for early laryngeal cancer diagnosis.


Sequence-driven telomeric chromatin structure.
Interstitial (also called internal or intrachromosomal) telomeric sequences (ITS) are found in many organisms.(1) In hamsters, CHO cells show long (up to several Mbp) ITS(2) (Fig. 1A) which are over involved in spontaneous or radiation induced chromosome aberrations.(3) ITS are also found in human, but they are much shorter (several hundreds of bp maximum) and show no radiosensitivity.(4) We report here the striking observation that interstitial telomeric chromatin, although located near centromeres in hamster cells, shares common structural features with truly telomeric chromatin (located at the end of chromosomes), and notably a higher nucleosome density than bulk chromatin, which correlates with a highly regular chromatin structure. This study emphasizes the critical role of DNA in establishing particular chromatin folding, confirming that the DNA sequence encodes important information concerning local chromatin properties.


Morphological and behavioral alterations in zebrafish larvae after exposure to contaminated river sediments collected in different weather conditions.
Wastewater treatment plants (WWTPs) are the primary source of micropollutants in aquatic ecosystems. Many micropollutants tend to bind to sediments and persist until remobilizion by bioturbation or flood events. Advanced effluent treatment by ozonation has been proven to eliminate most micropollutants. The present study characterizes sediments' toxic potential regarding zebrafish embryo development, which highly complex nervous system is vulnerable to exposure to neurotoxic substances. Furthermore, behavioral changes can be induced even at low pollutant concentrations and do not cause acute toxicity. The study area includes stretches of the main waterbody, the Wurm River (sampling sites W1-W5), and its tributary the Haarbach River (sampling sites H1, and H2) in North-Rhine Westphalia, Germany. Both waterbodies serve as recipients of WWTPs' effluents. The effluent entering the Haarbach River is conventionally treated, while the Wurm River receives ozonated effluent from the Aachen-Soers WWTP. Seven sampling sites up- and downstream of the WWTPs were investigated in June of two subsequent years. The first sampling campaign in 2017 was characterized by prolonged dry weather. The second sampling campaign in 2018 occurred after prolonged rain events and the release of the rainwater overflow basin. Direct exposure of zebrafish embryos to native sediments using the sediment contact test represented an ecologically realistic scenario and showed no acute sublethal effects. Exposure of the zebrafish embryo to freeze-dried sediments representing the ecotoxicological status of sediments during flood events unfolded acute sublethal toxicity. Behavioral studies with zebrafish larvae were an essential part of environmental neurotoxicity testing. Zebrafish larvae exposed to sediments' concentrations causing no acute effects led to behavioral changes signalizing neurotoxic substances in sediments. Polyaromatic hydrocarbons, polychlorinated biphenyls, and nitroaromatic compounds were identified as potential toxicity drivers, whereby the rainwater overflow basin served as a possible source of pollution. Mixture toxicity, effect-directed analysis, and further sediment monitoring are needed.


Empirical evidence for psychopharmacologic treatment in early-onset psychosis and schizophrenia.
Psychotic symptoms presenting in youth can be clinically complex and require that a child and adolescent psychiatrist use significant skill in making a diagnosis and initiating treatment. There are a number of illnesses to rule out before making a diagnosis of early-onset schizophrenia in particular. Psychosis in youth has significant associated morbidity and places high demands not only on families but also on the medical and educational systems. More effective pharmacologic and nonpharmacologic treatments for psychosis are needed. Nonpharmacologic therapies targeting relatively treatment-resistant domains of dysfunction such as neurocognition are also necessary as adjunctive treatments to our extant pharmacologic agents.


Decoding Human Megakaryocyte Development.
Despite our growing understanding of embryonic immune development, rare early megakaryocytes (MKs) remain relatively understudied. Here we used single-cell RNA sequencing of human MKs from embryonic yolk sac (YS) and fetal liver (FL) to characterize the transcriptome, cellular heterogeneity, and developmental trajectories of early megakaryopoiesis. In the YS and FL, we found heterogeneous MK subpopulations with distinct developmental routes and patterns of gene expression that could reflect early functional specialization. Intriguingly, we identified a subpopulation of CD42b+CD14+ MKs in vivo that exhibit high expression of genes associated with immune responses and can also be derived from human embryonic stem cells (hESCs) in vitro. Furthermore, we identified THBS1 as an early marker for MK-biased embryonic endothelial cells. Overall, we provide important insights and invaluable resources for dissection of the molecular and cellular programs underlying early human megakaryopoiesis.


Location of specific messenger RNAs in Caenorhabditis elegans by cytological hybridization.
We have developed an autoradiographic technique for detecting specific Caenorhabditis elegans messenger RNA molecules in situ by hybridization of labeled, cloned DNA probes to fixed tissue sections and squashes of embryos and adults. We report analyses with probes of actin and collagen gene sequences from a C. elegans genomic clone library. Hybridization is RNase sensitive and tissue specific. In adults the actin probe, which recognizes cytoplasmic as well as muscle actin mRNA, hybridizes strongly to muscle and distal gonad (ovary), somewhat less strongly to maturing oocytes, and weakly to intestine. The collagen probe hybridizes weakly to distal gonad and intestine and very strongly to subcuticular tissues, in particular to the hypodermal cells and syncytial cytoplasm of the lateral hypodermal ridges, which are the sites of cuticle synthesis. In embryos, hybridization to squashes indicates that actin message is present at fertilization, decreases during early cleavage, and then increases again during morphogenesis. By contrast, collagen message is absent until the 100-cell stage and then increases rapidly during morphogenesis. The number of cells labeled is consistent with the view that the collagen probe hybridizes to hypodermal precursor cells. We estimate that our present methods can detect messages representing about 0.2% or more of the total mRNA population, and increases in this sensitivity should be possible. Therefore, the cytological hybridization technique should be useful for determining temporal and spatial patterns of specific mRNA distributions during development, at least for abundant and moderately abundant messages.


Risk factors of cytomegalovirus infection after pediatric liver transplantation and effectiveness of preemptive therapy.
Cytomegalovirus (CMV) infection is the most common infection following pediatric liver transplantation (LT). Preemptive therapy (PET) is an approach to initiate antiviral treatment for asymptomatic early CMV viremia detected by surveillance testing. However, data on CMV infection after PET are scarce, and the optimal cut-off remains controversial. This study aimed to evaluate the incidence, risk factors, and consequences of CMV infection in pediatric LT using 2 different viral load (VL) cut-offs. We retrospectively reviewed patients aged 0-18 years who underwent LT at Ramathibodi Hospital between March 2001 and August 2020. Demographic data, CMV infection, CMV treatment, and consequences of CMV infection were collected. CMV viremia was monitored by a quantitative nucleic acid amplification test. Clinical outcomes were compared after starting antiviral therapy at a low (>400 but <2000 IU/mL) and a high VL cut-off (≥2000 IU/mL). A total of 126 patients were included. CMV infection was 71% (90/126), with an incidence rate of 5.5 per 1000 patient-day. Higher tacrolimus and prednisolone dosages were associated with CMV infection with an adjusted hazard ratio of 1.2 (95%CI 1.0-1.4, p = .02) and 2.4 (95%CI 1.9-3.4, p < .001), respectively. The consequences of CMV infection did not differ significantly for the low and high CMV VL cut-off groups. CMV infection in LT recipients is common and is associated with higher tacrolimus and corticosteroid dosage. Additionally, using the CMV VL cut-off at 2000 IU/mL to initiate antiviral therapy is practical and effective in preventing CMV disease.


Deep learning networks reflect cytoarchitectonic features used in brain mapping.
The distribution of neurons in the cortex (cytoarchitecture) differs between cortical areas and constitutes the basis for structural maps of the human brain. Deep learning approaches provide a promising alternative to overcome throughput limitations of currently used cytoarchitectonic mapping methods, but typically lack insight as to what extent they follow cytoarchitectonic principles. We therefore investigated in how far the internal structure of deep convolutional neural networks trained for cytoarchitectonic brain mapping reflect traditional cytoarchitectonic features, and compared them to features of the current grey level index (GLI) profile approach. The networks consisted of a 10-block deep convolutional architecture trained to segment the primary and secondary visual cortex. Filter activations of the networks served to analyse resemblances to traditional cytoarchitectonic features and comparisons to the GLI profile approach. Our analysis revealed resemblances to cellular, laminar- as well as cortical area related cytoarchitectonic features. The networks learned filter activations that reflect the distinct cytoarchitecture of the segmented cortical areas with special regard to their laminar organization and compared well to statistical criteria of the GLI profile approach. These results confirm an incorporation of relevant cytoarchitectonic features in the deep convolutional neural networks and mark them as a valid support for high-throughput cytoarchitectonic mapping workflows.


Statistical Process Control (SPC) to drive improvement in length of stay after colorectal surgery.
Novel quality improvement(QI) methods are needed to optimize healthcare costs and value. Our goal was to determine if Statistical Process Control(SPC), an industrial QI tool, could transform length of stay(LOS) into a process measure, identify outliers, and their impact on surgical outcomes. SPC was performed on an institutional colorectal resection database 1/1/13-5/1/2018 to identify outliers and compare outcome variables across outliers and non-outliers. Control charts analyzed the process performance of LOS over time. Control limits were set at ± 1 standard deviation(SD) from the mean. Measures were stable within these limits. LOS was stable, with consistent annual rates and variation of outliers. Outliers had identifiable causes of variation that were significantly different from non-outliers(p < 0.05). The variation resulted in more complications, readmissions, and reoperations in outliers(p < 0.05). SPC can be applied to LOS, a stable process measure with decreasing variability over time, and easy outlier identification. Identifying outliers can facilitate targeted quality improvement.


Topical application of Polygonum multiflorum extract induces hair growth of resting hair follicles through upregulating Shh and β-catenin expression in C57BL/6 mice.
Polygonum multiflorum has traditionally been used for treating patients suffering from baldness and hair loss in East Asia. The present study sought to investigate the hair growth promoting activities of Polygonum multiflorum and its mechanism of action. The Polygonum multiflorum extract was topically applied to the shaved dorsal skin of telogenic C57BL6/N mice. To determine the effect of Polygonum multiflorum extract in telogen to anagen transition, the expression of β-catenin and Sonic hedgehog (Shh) was determined by immunohistochemistry analysis. Polygonum multiflorum extract promoted hair growth by inducing anagen phase in telogenic C57BL6/N mice. In Polygonum multiflorum extract treated group, we observed increase in the number and the size of hair follicles that are considered as evidence for anagen phase induction. Immunohistochemical analysis revealed that earlier induction of β-catenin and Shh were observed in Polygonum multiflorum extract treated group compared to that in control group. These results suggest that Polygonum multiflorum extract promotes hair growth by inducing anagen phase in resting hair follicles.


Effects of a hospital-based leisure activities programme on nurses' stress, self-perceived anxiety and depression: A mixed methods study.
To determine the effects of a hospital-based leisure activities programme on nurses' stress, self-perceived anxiety and depression. Nursing work in clinical settings is highly stressful and may result in an increase in nurses' turnover rate, which threatens the quality of nursing care and patient safety. We used a mixed methods design and a three-month intervention (January to April, 2019) involving a convenience sample of 176 nurses working at a Chinese tertiary hospital. We conducted 12 semi-structured interviews and performed a content analysis. The pre- and post-intervention comparisons of nurses' stress, self-perceived anxiety and depression were performed using a paired t test. The 3-month leisure activities programme significantly decreased nurses' job stress (t = 3.80, p < .01), perceived personal stress (t = 3.30, p < .01), self-perceived anxiety (t = 3.76, p < .01) and depression (t = 2.73, p < .01). The qualitative findings revealed five mechanisms linking leisure activities to subjective well-being: detachment recovery, autonomy, mastery, meaning and affiliation. A hospital-based leisure activities programme had a positive effect on job stress, self-perceived anxiety and depression, thus improving nurses' well-being. A hospital-based leisure activities programme provides a beneficial strategy for ameliorating nurses' psychosocial issues. Interventions aimed at facilitating or increasing nurses' participation in leisure activities are greatly needed.


Fertility preservation knowledge, attitudes and intentions among children by proxy and adolescents with cancer.
The current study aimed to assess knowledge, attitudes and intentions in relation to fertility preservation among children, by proxy, and adolescents with cancer. Although fertility preservation options have been developed to mitigate the adverse long-term effects of life-saving cancer treatment on fertility, fertility is difficult for children and adolescents to conceptualize, especially when they face a cancer diagnosis. This was a descriptive, semi-quantitative analysis. Adolescents and parents of children or adolescents within 6 months of a cancer diagnosis and undergoing gonadotoxic treatment were invited to participate. Seventy-one families completed and returned the study's questionnaire (91 questionnaires) over a period of 26 months. The vast majority of participants were aware of the gonadotoxic effects of cancer treatment (85%) and had positive attitudes towards fertility preservation (>90%), but only a portion of them (20%) were willing to take action towards this goal. Although adolescent-parent pairs had similar attitudes towards fertility preservation, adolescents tended to be more sceptical about experimental fertility preservation options. Male post-pubertal cancer patients were more likely to be offered fertility preservation counselling or referral to a specialist in comparison to their younger and female counterparts. Fertility preservation care has advanced but there are still gender and age differences in counselling and treatment initiation in the paediatric and adolescent cancer population. Interventions to improve provider-patient-parent communication regarding fertility preservation and to help patients address the observed intention-behaviour gap in relation to fertility preservation options are needed.


Use of a proline-specific endopeptidase to reintroduce gluten in patients with non-coeliac gluten sensitivity: A randomized trial.
A gluten-free diet (GFD) is the main therapy for non-coeliac gluten sensitivity (NCGS). However, the availability of novel enzymes with the ability to digest gluten could represent a therapeutic opportunity for NCGS patients to avoid a GFD. To evaluate the controlled reintroduction of gluten with or without the endopeptidase P1016 in NCGS patients. This is a randomized, double-blind, placebo-controlled monocentric study, Registered under ClinicalTrials.gov Identifier no. NCT01864993. Gluten was reintroduced incrementally over a 3-week period under nutritional control. NCGS patients were randomized into two groups and administered P1016 or placebo during gluten reintroduction. We evaluated symptoms (visual analogue scale, VAS), quality of life (SF-36) and mental health symptoms (SCL-90) on a weekly basis. We enrolled a total 23 patients who were allocated to a placebo group (n = 11, age 38.4 ± 2.9) or an intervention group (n = 12, age 39.5 ± 3.1). No effect of P1016 on symptoms was found. During gluten reintroduction, patients reported a significant increase in abdominal pain and a worsening of stool consistency. Furthermore, no differences were found between the groups regarding SCL-90 and SF-36 scores. Our results demonstrate a lack of effect of P1016 in the management of NCGS patients and the possible reintroduction of gluten.


Does immunosuppressive treatment ameliorate morphology changes in encapsulating peritoneal sclerosis?
Encapsulating peritoneal sclerosis (EPS) is a clinical syndrome associated with ileus symptoms and irreversible sclerosis of the peritoneal membrane. Inflammation, fibrosis, and neoangiogenesis are the main features of the pathophysiology. No evidence-based therapy is currently available for EPS. In recent years, anti-inflammatory and immunosuppressive (IS) treatment modalities have become more popular. The aim of the present study was to investigate the effects of various IS treatment strategies-glucocorticosteroid (GC), azathiopurine (AZT), and cyclosporin (CsA)- on regression of EPS. We divided 52 nonuremic Wistar albino rats into six groups: Control group-2 mL isotonic saline injected intraperitoneally (IP) daily for 3 weeks; CG group-2 mL/200 g 0.1% chlorhexidine gluconate (CG) and 15% ethanol dissolved in saline injected IP daily for 3 weeks; Resting group-CG (weeks 1 - 3), plus peritoneal rest (weeks 4 - 6); Corticosteroid (GC) group-CG (weeks 1 - 3), plus 10 mg/L prednisolone in drinking water (weeks 4 - 6); AZT group- CG (weeks 1 - 3), plus 100 mg/L azathioprine in drinking water (weeks 4 - 6); and CsA group-CG (weeks 1 - 3), plus cyclosporin 7.5 mg/kg by subcutaneous injection daily (weeks 4 - 6). At the end of the study, under ketamine HCl anesthesia, the rats were humanely killed by bleeding. Parietal peritoneal samples were then taken from same location (away from the injection site) and changes of parietal peritoneum morphology were examined by a single pathologist. The CG severely disturbed parameters of peritoneal morphology, increasing peritoneal thickness, inflammatory activity, vascularity, and fibrosis score as compared with the Control group (p < 0.05). No benefit was observed for any parameter in the Resting group as compared withthose parameters in the CG group (p < 0.05). We observed a lower fibrosis score and less peritoneal thickness in the GC group as compared with the Resting group (p < 0.05). No beneficial effects of AZT on peritoneal morphology were observed as compared with the effects of peritoneal rest or corticosteroid therapy. Treatment with cyclosporin resulted in more fibrosis, vascularity, and inflammation than was seen with corticosteroid therapy (p < 0.05). Immunosuppressive therapies, especially those that are corticosteroid-based, may have therapeutic value in the management of EPS. Patients treated with cyclosporin may have a risk for developing EPS.


Long-term effect of total fundoplication on the myotomized esophagus.
From 1978 to 1983, 17 patients had an esophagocardiomyotomy with an added short total fundoplication as an antireflux procedure. Thirteen had achalasia and 4, diffuse esophageal spasm. All patients initially had the usual symptoms of these motor disorders. Early after the operation all became asymptomatic, but over the years of follow-up, symptoms reappeared in 14 of 17 patients, and 5 required reoperation. The distal esophageal transverse diameter showed progressive dilatation from 3.9 cm preoperatively to more than 6 cm after 10 years of evolution. Over the same period, deterioration in the esophageal emptying capacity caused esophageal stasis to increase from 32% to 75%. Manometric changes were significant after the operation: resting pressures in the esophageal body decreased from 10.5 to 4.4 mm Hg (p < 0.001) proximally and from 12.2 to 4.6 mm Hg distally (p < 0.001). Peak contraction pressures became significantly weaker: 38 to 30 mm Hg in the proximal esophagus (p < 0.001) and from 49.2 to 28.1 in the distal esophagus (p < 0.001). Tertiary contractions were unchanged distally, but peristalsis reappeared in more than 30% of all swallows in the proximal half of the esophageal body. The resting pressure gradient in the lower esophageal sphincter area was reduced from 25.5 to 7.4 mm Hg by the operation. This gradient remained stable over 10 years of follow-up. No significant acid exposure was documented in 8 patients undergoing 24-hour pH recordings after their operation. Endoscopy revealed dilatation and retention without evidence of reflux esophagitis damage. Total fundoplication when associated with esophageal myotomy results in improved symptoms in the early postoperative phase.(ABSTRACT TRUNCATED AT 250 WORDS)


Preformed antibodies to alpha1,3-galactosyltransferase gene-knockout (GT-KO) pig cells in humans, baboons, and monkeys: implications for xenotransplantation.
The aim of our study was to determine the prevalence and cytotoxicity of primate antibodies directed to antigens other than Galalpha1,3Gal (Gal), termed nonGal antigens. Sera from human, baboon, and cynomolgus monkeys were tested by flow cytometry for IgM and IgG binding to both wild-type (WT) and GT-KO pig peripheral mononuclear cells (PBMC). Also, complement-dependent cytotoxicity assays were performed. All species demonstrated significantly higher antibody binding and cytotoxicity to WT cells compared to GT-KO cells (P < .01). Cynomolgus monkeys had significantly higher IgM binding to WT and GT-KO cells than did baboons or humans (P < .01). Furthermore, approximately 50% of both human and baboon sera proved to be lytic to GT-KO cells, compared to 76% of monkey sera (P < .01). We confirm the advantage of using GT-KO pig grafts over WT pig grafts. However, our results suggest that, compared to the cynomolgus monkey, the baboon may be a more suitable model to study antibody-mediated rejection of GT-KO pig grafts.


Implications from COVID-19 for future pandemic global health governance.
Limitations of current global health governance revealed during the COVID-19 pandemic can inform the ongoing deliberations of an international treaty on pandemics. To report on WHO definitions for governance and the enforcement of treaties in the context of a proposed international treaty on pandemics. This narrative review was based on keyword searches related to public health, global health governance, and enforcement in PubMed/Medline and Google Scholar. Snowballing for additional articles followed the keyword search review. WHO lacks a consistent definition of global health governance. Moreover, in its current state, the proposed international treaty on pandemics lacks articulated compliance, accountability, or enforcement mechanisms. Findings reveal that humanitarian treaties often fail to achieve their aims absent clear enforcement mechanisms. The proposed international treaty on public health is garnering a range of perspectives. Decision-makers should evaluate whether a globally aligned definition of global health governance is needed. Decision-makers should also consider whether the proposed international treaty on pandemics should be opposed if it lacks sufficiently clear compliance, accountability, and enforcement mechanisms. To our knowledge, this narrative review is believed to be the first of its kind to search scientific-oriented databases regarding governance and international pandemic treaties. The review includes several findings that advance the literature. These findings, in turn, reveal two key implications for decision-makers. First, whether an aligned definition for governance addressing compliance, accountability, and enforcement mechanisms is needed. Second, whether a draft treaty lacking enforcement mechanisms should be approved.


Mdivi-1 induces spindle abnormalities and augments taxol cytotoxicity in MDA-MB-231 cells.
Taxol is a first-line chemotherapeutic for numerous cancers, including the highly refractory triple-negative breast cancer (TNBC). However, it is often associated with toxic side effects and chemoresistance in breast cancer patients, which greatly limits the clinical utility of the drug. Hence, compounds that act in concert with taxol to promote cytotoxicity may be useful to improve the efficacy of taxol-based chemotherapy. In this study, we demonstrated that mdivi-1, a putative inhibitor of mitochondrial fission protein Drp1, enhances the anticancer effects of taxol and overcomes taxol resistance in a TNBC cell line (MDA-MB-231). Not only did mdivi-1 induce mitotic spindle abnormalities and mitotic arrest when used alone, but it also enhanced taxol-induced antimitotic effects when applied in combination. In addition, mdivi-1 induced pronounced spindle abnormalities and cytotoxicity in a taxol-resistant cell line, indicating that it can overcome taxol resistance. Notably, the antimitotic effects of mdivi-1 were not accompanied by prominent morphological or functional alterations in mitochondria and were Drp1-independent. Instead, mdivi-1 exhibited affinity to tubulin at μM level, inhibited tubulin polymerization, and immediately disrupted spindle assembly when cells entered mitosis. Together, our results show that mdivi-1 associates with tubulin and impedes tubulin polymerization, actions which may underlie its antimitotic activity and its ability to enhance taxol cytotoxicity and overcome taxol resistance in MDA-MB-231 cells. Furthermore, our data imply a possibility that mdivi-1 could be useful to improve the therapeutic efficacy of taxol in breast cancer.


Second- and third-trimester therapeutic terminations of pregnancy in cases with complete placenta previa--does feticide decrease postdelivery maternal hemorrhage?
To study the feasibility of second- and third-trimester termination of pregnancy (TOP) with complete placenta previa, and the impact of performing feticide before labor induction on maternal hemorrhagic morbidity. From 1987 to 2002, the databases of two referral hospitals were reviewed. We identified 15 cases of second- or third-trimester TOP in women with complete placenta previa. Feticide was performed 2-14 days before induction in 6/15 cases. Cervical ripening was achieved in 8 cases by mifepristone alone (n = 2) or by mifepristone and dilapan (n = 6). Labor was induced by vaginal gemeprost (n = 2), intramuscular (n = 5) or intravenous (n = 4) sulprostone, vaginal misoprostol (n = 1) or a combination of misoprostol and sulprostone (n = 3). Hemorrhage was defined by the need for transfusion. The difference between the preoperative and the lowest per- or postoperative maternal hemoglobin level was also analyzed. Of the 9 women who underwent labor induction without previous feticide, 4 required blood transfusions, 1 of whom had a hemostat hysterectomy. The mean hemoglobin difference was 2.5 g/dl (range: 0.5-5.3). None of the 6 patients with preinduction feticide required transfusion. The hemoglobin difference was significantly smaller in this group than in terminations without previous feticide (mean: 1.0 g/dl ; range: 0.1-2.2; p = 0.03). In cases with complete placenta previa, second- or third-trimester TOP is feasible. It carries a substantial risk of hemorrhage that might be decreased by preinduction feticide.


Massive acetaminophen overdose with metabolic acidosis refractory to N-acetylcysteine, fomepizole, and renal replacement therapy.
Massive Acetaminophen (N-acetyl-p-aminophenol; APAP) overdose is a common presentation to emergency departments around the world. While N-acetylcysteine (NAC) remains the cornerstone of treatment for APAP overdose, extracorporeal treatment, in the form of renal replacement therapy with intermittent hemodialysis (IHD) or continuous renal replacement therapy (CRRT) may provide benefit in cases associated with altered mental status and metabolic acidosis. One treatment with IHD is typically sufficient for resolution of acidosis and global improvement clinically. We describe a case of massive APAP overdose presenting with altered mental status and lactic acidosis, refractory to multiple treatments of IHD as well as CRRT and high-dose NAC along with fomepizole. Despite these interventions, fulminant liver failure progressed with cerebral edema, coagulopathy and death. This is the first description of a fatal acetaminophen ingestion refractory to both IHD and prolonged CRRT. This case highlights the need for further investigation in the management of massive APAP overdose, including optimal method and timing of renal replacement therapy.


Body Composition Changes Following a Very-Low-Calorie Pre-Operative Diet in Patients Undergoing Bariatric Surgery.
Fatty liver in obese patients increases the technical difficulty of bariatric surgery. Pre-operative weight loss with a very-low-calorie diet (VLCD) is commonly used to facilitate surgery. Few studies have quantified the systemic effect of rapid pre-operative weight loss on body composition. The objective of this study is to evaluate body composition changes in bariatric surgery patients undergoing a VLCD. Body composition assessments were performed between August 2017 and January 2019 using dual-energy X-ray absorptiometry immediately before and after a 2-week VLCD at St Vincent's Hospital Melbourne. Data collected prospectively pre- and post-VLCD included total body weight, excess body weight, body mass index (BMI), lean body mass (LBM), fat mass (FM) and bone mineral content (BMC). The pre- and post-operative results were compared. Forty-four patients completed both the 2-week VLCD and body composition assessments. Following a 2-week VLCD, patients lost a mean of 4.5 kg (range - 0.3 to 9.5) in a total body weight and 8.8% (range - 0.9 to 17.1) of excess body weight, with a mean reduction in body mass index of 1.6 kg/m2 (range - 0.2 to 3.1). Loss of LBM was 2.8 kg and was significantly greater than loss of FM, 1.7 kg (p < 0.05). BMC changes were insignificant. A VLCD is an effective tool for pre-operative weight reduction. In this cohort, a large amount of the total weight loss was attributed to a loss of lean body mass. The impact of significant lean body mass loss and its relationship to short- and long-term health outcomes warrants further assessment.


Factoring in the spatial effects of symbolic number representation.
Spatial constituents of adult symbolic number representation produce effects of size-value congruity, Spatial Numerical Association of Response Codes (SNARC), and numerical distance. According to behavioral experiments, these effects belong to distinct processing stages. Yet, these effects evoke overlapping responses in both early and late Event Related Potentials (ERPs). To probe whether these overlaps indicate sharing of resources, all relevant stimulus and response conditions were factorially combined in a numerical value comparison task. To secure ERP validity, same numbers were compared against variable reference values. This design resulted in previously unobserved interactions in behavior but inhibited late ERP effects. All effects arose early in the P1 component (around 100 ms) and most showed hemispheric specificity. Independency of congruity and SNARC effects was observed, whereas SNARC and numerical distance were closely intertwined. Differences in hemispheric specificity, rather than stage-wise separation, were key to independence.


Arginine/asymmetric dimethylarginine ratio and cardiovascular risk factors in patients with predialytic chronic kidney disease.
Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase (NOS) that accumulates in patients with chronic kidney disease (CKD) and predicts cardiovascular outcome. Arginine is the substrate for NOS and a low arginine/ADMA ratio may lead to a reduced NO production and a worse prognosis. We have studied how other important CKD variables predict the arginine/ADMA ratio. The population is 160 predialytic CKD patients (median age 61 years). We used backward stepwise regression to identify the best predictors of p-arginine, p-ADMA and arginine/ADMA ratio. P-ADMA was predicted by estimated GFR (eGFR) (adjusted R(2)=0.17, p>0.00). Arginine/ADMA ratio was predicted by gender, eGFR, use of renin angiotensin aldosteron (RAAS) inhibitors, current smoking and use of platelet inhibitors (adjusted R(2)=0.18, p<0.00). Reduced eGFR is associated with reduced arginine/ADMA ratio. The use of RAAS inhibitors and male gender may be protective against a low arginine/ADMA ratio.


Tobacco use among married women in Nepal: the role of women's empowerment.
This study documented the prevalence and correlates of tobacco use among women of reproductive age in Nepal using nationally representative data. We utilized the 2006 Nepal Demographic and Health Survey that interviewed 10,793 women and 4,397 men. We analyzed the couple's data or households (N = 2,600) in which both husband and wife were interviewed. We examined the effects of women's empowerment-measured by education, employment, intra-household decisions, and age-on their tobacco use controlling for other individual and household characteristics. Women's empowerment had mixed effects on tobacco use. While women's education was inversely associated with their tobacco use, their age, employment and ability to make intra-household mobility decisions were positively associated with smoking. Women with primary and beyond primary education were 48 and 92 % less likely to smoke compared to women with no education, respectively. Tobacco use among women increased dramatically with age from 8 % in teen years to 42 % in their forties. A 1 year increase in age increased the odds of tobacco use by 6 %. Women whose husbands smoked were twice as likely to smoke. Nepal should not only restrict tobacco use in public places by implementing its Tobacco Control and Regulatory Act of 2010 but also focus on encouraging smoke-free homes by increasing awareness about the health consequences of tobacco use and secondhand smoke among populations most likely to smoke that include nearly all men, employed women, women with low levels of education, women whose spouses smoke and those who are 30 and above in age. Additionally, a long term goal should be to ensure at least 5th grade of education for all girls.


Management options of non-syndromic sagittal craniosynostosis.
There have been various effective surgical procedures for the treatment of non-syndromic sagittal craniosynostosis, but no definitive guidelines for management have been established. We conducted a study to elucidate the current state of practice and establish a warranted standard of care. An Internet-based study was sent to 180 pediatric neurosurgeons across the country and 102 craniofacial plastic surgeons in fourteen different countries, to collect data for primary indication for surgical management, preference for timing and choice of surgery, and pre-, peri-, and post-operative management options. The overall response rate from both groups was 32% (n=90/284). Skull deformity was the primary indication for surgical treatment in patients without signs of hydrocephalus for both neurosurgeons and craniofacial surgeons (80% and 63%, respectively). Open surgical management was most commonly performed at six months of age by neurosurgeons (46%) and also by craniofacial surgeons (35%). Open surgical approach was favored for patients younger than four months of age by neurosurgeons (50%), but endoscopic approach was favored by craniofacial surgeons (35%). When performing an open surgical intervention, most neurosurgeons preferred pi or reversed pi procedure (27%), whereas total cranial vault remodeling was the most commonly performed procedure by craniofacial surgeons (37%). The data demonstrated a discrepancy in the treatment options for non-syndromic sagittal craniosynostosis. By conducting/comparing a wide survey to collect consolidative data from both groups of pediatric neurosurgeons and craniofacial plastic surgeons, we can attempt to facilitate the establishment of standard of care.


Long-Lived Trees Are Not Immortal.
Separating out the different effects of ageing on long-lived trees remains challenging. Herein current approaches used to explore senescence in millennial trees are highlighted. Molecular and biochemical analyses of the vascular cambium provide novel insight into the extent to which millennial trees can withstand the wear and tear of ageing.


Vitamin E and immunity.
Vitamin E is the most important chain-breaking, lipid-soluble antioxidant present in body tissues of all cells and is considered the first line of defense against lipid peroxidation and it is important for normal function of the immune cells. However, vitamin E deficiency is rare in well-nourished healthy subjects and is not a problem, even among people living on relatively poor diets, both T- and B-cell functions are impaired by vitamin E deficiency. While immune cells are particularly enriched in vitamin E because of their high polyunsaturated fatty acid content, this point puts them at especially high risk for oxidative damage. Besides its immunomodulatory effects, vitamin E also plays an important role in carcinogenesis with its antioxidant properties against cancer, and ischemic heart disease with limiting the progression of atherosclerosis. Supplementation of vitamin E significantly enhances both cell mediated and humoral immune functions in humans, especially in the elderly and animals.


Iodine nutrition: Disorders, monitoring and policies.
Iodine is an essential mineral nutrient and an integral component of thyroid hormones. Iodine deficiency is typically associated with goiter, but can have more serious health implications. Adequate iodine status is important for normal brain development. Iodine deficiency in utero or in early life can cause severe neurological and cognitive impairment. Over the last three decades, global efforts have reduced the prevalence of iodine deficiency disorders (IDD) in many areas of the world with implementation of nutrition policies and programs such as "salt" iodization. However, in a number of areas iodine deficiency is still widespread. Iodine deficiency in remote regions with high poverty will be more difficult to eradicate. Efforts to eliminate IDD in affected areas and sustaining successful iodine programs will be a priority given the substantial public health and economic benefits. A key component will be periodic monitoring of population iodine status to ensure sufficient intakes and the absence of excessive intakes. Median urinary iodine concentration (UIC), a validated biomarker for assessing population iodine status, will facilitate monitoring. Research validating "usual" UIC for use in combination with the Estimated Average Requirement cut-point method will expand its utility and allow accurate determination of the prevalence of inadequate intakes in populations. Further research on the development of biomarkers for assessment of individual iodine status for routine patient care will be important.


Beyond Intraocular Pressure: Visual Functioning and Quality of Life in Primary Congenital Glaucoma and Secondary Childhood Glaucoma.
To compare the visual functioning (VF) and vision-related QoL (VRQoL) of children 8-18 years old treated for primary congenital glaucoma (PCG) and secondary childhood glaucoma. Cross-sectional study. A total of 309 children 8-18 years old treated for PCG and secondary childhood glaucoma between 2000 and 2010 by a single pediatric glaucoma specialist were prospectively enrolled at LV Prasad Eye Institute, Hyderabad, India. Children completed 2 questionnaires, the LV Prasad Functional Vision Questionnaire-II (LVP-FVQ-II), and the Impact of Vision Impairment-Children (IVI-C) questionnaire. Rasch-calibrated scores from both these questionnaires were used to compare the VF and VFQoL between the 2 groups. Mean ages of the children were 12.2 and 12.6 years in the PCG (53%, median age at diagnosis = 5 months) and secondary glaucoma groups (47%, median age at diagnosis = 3 years), respectively. A majority (80%) of children had bilateral glaucoma and underwent filtering surgery (83%). Mean better eye logMAR visual acuity (VA) was comparable between PCG and secondary childhood glaucoma groups (0.49 vs 0.52, respectively; P = 0.59). Children with PCG reported significantly better VF and VRQoL than secondary childhood glaucoma patients. Unadjusted and adjusted childhood glaucoma group comparisons revealed secondary childhood glaucoma to be associated with worse VF and VRQoL compared to PCG (difference for VF, -0.83; 95% confidence interval [CI], -1.34 to 0.31; P = 0.002; 0.39; 95% CI, 0.16-0.62; P = 0.001 for VRQoL). Results show that children with treated PCG experience significantly better VF and VRQoL than those with secondary childhood glaucoma, despite comparable VA and IOP.


The association between motor modules and movement primitives of gait: A muscle and kinematic synergy study.
In spite of the extensive literature on the analysis of the muscle synergies during gait, the functionality of these synergies has not been studied in detail. This study explored the relationship between the motor modules and the kinematic maneuvers involved in human walking. Motion and surface electromyography data (of 28 trunk and lower extremity muscles) were acquired from ten healthy subjects during ten trials of self-selected speed gait each. The joint angle trajectories were half-wave rectified and divided into two independent positive directional degrees-of-freedom. The muscle and kinematic synergies were both extracted using the non-negative matrix factorization (NNMF) technique and clustered via k-means method. Results indicated that for both the muscle and kinematic synergies, a same number of modules (five) could reconstruct the 200 limb-trial data reasonably well. Moreover, each individual muscle synergy was found to be associated with a single kinematic synergy, based on the similar activation periods of the paired muscle and kinematic modules and the high correlation of their activation patterns (r = 0.88 ± 0.05), with a consistent phase advance for muscle synergies (mean = 7.59 ± 2.34 %). It was concluded that there is a one-to-one association between the motor modules and kinematic synergies, suggesting that each individual kinematic synergy, representing a movement primitive of human walking, might be implemented by recruitment of a paired motor module.


Thermodynamics of oxidative phosphorylation in bovine heart submitochondrial particles.
The rates of both forward and reverse electron transfer in phosphorylating submitochondrial particles from bovine heart can be controlled by the thermodynamic phosphorylation potential (deltaGp) of the adenine nucleotide system. deltaGp is the Gibbs free energy of ATP synthesis and is defined by the relationship deltaGp = -deltaG'o + RTln([ATP]/[ADP][Pi]) where deltaG'o is the standard free energy of ATP hydrolysis. Studies of the effects of deltaGp on NADH respiration and the reduction of NAD+ by succinate show that increasing values of deltaGp cause an inhibition of forward electron transfer and a stimulation of reverse electron transfer. Between deltaGp values of 7.6 and 13.0 kcal/mol the rate of NADH respiration decreased 3-fold and the rate of NAD+ reduction by succinate increased 3-fold. Indirect phosphorylation potential titration experiments as well as direct chemical measurements indicate that steady state levels of ATP, ADP, and Pi are established during NADH respiration which correspond to a deltaGp equal to 10.7 to 11.4 kcal/mol.


Contribution of human esterases to the metabolism of selected drugs of abuse.
Human esterases such as the human carboxylesterases (hCES) are important for the catalytic ester hydrolysis of xenobiotics and they play an important role in the detoxification of drugs (e.g., cocaine) but also in the activation of prodrugs (e.g., ramipril). Therefore, the aim of the presented study was to characterize the enzyme-catalyzed ester hydrolysis of ten drugs (cocaine, dimethocaine, ethylphenidate, 4-fluoro-3α-tropacocaine, 4-fluoro-3β-tropacocaine, heroin, methylphenidate, mitragynine, ramipril, and thebacon) by different esterase-containing systems (recombinant hCES1b, hCES1c, and hCES2, pooled human liver microsomes, pooled human liver S9 fraction, and pooled human plasma). Michaelis-Menten kinetic studies were done using in vitro incubations with the aforementioned enzyme-containing systems and LC coupled to ion trap MS for analysis. Ramipril and heroin were used as known model substrates to ensure reliable incubation conditions. The hydrolysis reactions followed classic Michaelis-Menten kinetics with exception of cocaine and 4-fluoro-3α-tropacocaine, for which hydrolysis rate was too low for reliable modeling. The substrates were mainly metabolized by the following enzymes systems: cocaine, hCES1c; dimethocaine, human plasma esterases; ethylphenidate, hCES1c; 4-fluoro-3β-tropacocaine, human plasma esterases; heroin, hCES2; methylphenidate, hCES1c; mitragynine, hCES1c; ramipril, hCES1b; thebacon, hCES2. Compounds bearing a small alcohol part and a larger acyl part showed higher affinity to hCES1 while those with a large alcohol part showed higher affinity to hCES2. The collected data are important for prediction of drug-drug or drug-food interactions as well as for individual variations in metabolism of drugs of abuse due to enzyme polymorphisms.


Clinical outcomes and cost-effectiveness of large-scale midwifery-led, paediatrician-overseen home phototherapy and neonatal jaundice surveillance: A retrospective cohort study.
To evaluate a large midwifery-led, paediatrician-overseen home jaundice surveillance and home phototherapy (HPT) programme. We conducted a retrospective cohort study over 2019. Included were all infants with birth gestation ≥35 weeks, discharged at 4-96 h and receiving care from midwifery-at-home (a 12-h daily, 365-days hospital-based outreach service, supported by hospital paediatricians). Phototherapy was delivered via BiliSoft blanket with treatment thresholds determined by standard nomograms. The main outcomes of interest were unplanned readmissions, and cost-effectiveness based on hospital finance department actual costs. Also examined were parental compliance, device issues and safety. During 2019, 4308 infants received home jaundice surveillance with 86% hospital-discharged before 72 h, 82% exclusively breastfed and 69% having overseas-born mothers. Four hundred infants received HPT, comprising 101 continuing from inpatient phototherapy (IPT), 56 rebounding after IPT, and 243 home-diagnosed as needing phototherapy and triaged to HPT. Only 1 of 400 (0.25%) HPT infants required readmission. Additionally, there were 80 home-diagnosed jaundiced infants triaged to immediate readmission for IPT. Maximal serum bilirubin was 454 μmol/L. No exchange transfusion, encephalopathy or HPT-device problems occurred. An early 2019 bilirubin analyser upgrade resulted in higher bilirubin readings and some unintended subthreshold phototherapy. Supported by midwives, most parents managed HPT with ease. HPT cost $640/day compared to $2100/day for infant IPT readmission and $1000/day for a longer birth-admission stay. Up to 2 weeks' midwifery-at-home care for the whole cohort cost $2 m less than a 2-day longer birth-admission stay. Large-scale, midwifery-led, paediatrician-overseen jaundice surveillance and HPT can achieve very low unplanned readmission rates and be cost-effective.


1-(4-Chloro-phen-yl)-3-{5-[(E)-2-phenyl-ethen-yl]-1,3,4-thia-diazol-2-yl}urea.
In the title compound, C(17)H(13)ClN(4)OS, the 1,3,4-thia-diazole ring makes dihedral angles of 9.70 (15) and 7.22 (10)° with the benzene and phenyl rings, respectively; the dihedral angle between these two rings is 6.37 (19)°. In the crystal, pairs of N-H⋯N and C-H⋯O hydrogen bonds between inversion-related mol-ecules result in supra-molecular ribbons displaying alternate R(2) (2)(8) and R(2) (2)(14) graph-set ring motifs.


Computational and spectral studies of 6-phenylazo-3-(p-tolyl)-2H-chromen-2-one.
6-Phenylazo-3-(p-tolyl)-2H-chromen-2-one 4 was prepared and characterized by IR, (1)H, and (13)C NMR spectral studies. The optimized structure of the chromen-2-one 4 was investigated by the Gaussian 03 B3LYP density functional method calculations at 6-31G(d,p) basis set. The gauge-independent atomic orbital (GIAO) (13)C and (1)H chemical shift calculations for the synthesized chromen-2-one in CDCl3 were also made by the same method. The computed IR frequencies of the chromen-2-one and the corresponding vibrational assignments were analyzed by means of potential energy distribution (PED%) calculation using vibrational energy distribution analysis (VEDA) program. The first order hyperpolarizability (βtot), polarizability (α) and dipole moment (μ) were calculated using 6-311G(d,p) basis set and the nonlinear optical (NLO) properties are also addressed theoretically. Stability of the chromen-2-one 4 molecule has been analyzed by calculating the intramolecular charge transfer using natural bond order (NBO) analysis. The molecular electrostatic potentials, HOMO-LUMO energy gap and geometrical parameters were also computed. Topological properties of the electronic charge density in chromen-2-one 4 were analyzed employing the Bader's Atoms in Molecule (AIM) theory which indicated the presence of intramolecular hydrogen bond in the molecule.


Behavior therapy for irritable bowel syndrome. A case report.
The behavior analysis and treatment of a patient with chronic irritable bowel syndrome are presented. The usefulness of this approach in a patient with a poor treatment prognosis is discussed.


Development of interdigitated arrays coated with functional polyaniline/MWCNT for electrochemical biodetection: application for human papilloma virus.
In this study, polyaniline-multiwalled carbon nanotube film (PANi-MWCNT) has been polymerized on interdigitated platinum electrode arrays (IDA), fabricated by MEMS technology for the detection of human papillomavirus (HPV) infection, using immobilized peptide aptamers as affinity capture reagent. Label-free, electrochemical detection of the specific immune reaction between antigen peptide aptamer HPV-16-L1 (with a molecular weight of 1825 Da), the most common genotype in cytological normal women worldwide, and its specific antibody of HPV-16 (which is much bigger with molecular weight of ca. 150 kDa) on multifunctional PANi-MWCNT based arrays was reported. The most significant advantage of this technique consists of reagentless and multiple detection of antigen-antibody complex formation on well conducting IDA interface of PANi-MWCNT, without intermediate steps or any labeling reagents, as normally required in the previous works.


Decoration of Squalenoyl-Gemcitabine Nanoparticles with Squalenyl-Hydroxybisphosphonate for the Treatment of Bone Tumors.
Therapeutic perspectives of bone tumors such as osteosarcoma remain restricted due to the inefficacy of current treatments. We propose here the construction of a novel anticancer squalene-based nanomedicine with bone affinity and retention capacity. A squalenyl-hydroxybisphosphonate molecule was synthetized by chemical conjugation of a 1-hydroxyl-1,1-bisphosphonate moiety to the squalene chain. This amphiphilic compound was inserted onto squalenoyl-gemcitabine nanoparticles using the nanoprecipitation method. The co-assembly led to nanoconstructs of 75 nm, with different morphology and colloidal properties. The presence of squalenyl-hydroxybisphosphonate enhanced the nanoparticles binding affinity for hydroxyapatite, a mineral present in the bone. Moreover, the in vitro anticancer activity was preserved when tested in commercial and patient-treated derived pediatric osteosarcoma cells. Further in vivo studies will shed light on the potential of these nanomedicines for the treatment of bone sarcomas.


Implications of the metabolic syndrome: the new epidemic.
On the basis of traditional risk factors, a large number of individuals in the United States can be classified as at intermediate risk for the development of ischemic heart disease. The diagnosis of the metabolic syndrome can help determine whether patients at intermediate risk should be considered for more aggressive risk-factor reduction. The measurement of novel risk factors, such as inflammatory markers, can identify a group of patients at high intermediate risk. The Adult Treatment Panel of the National Cholesterol Education Program suggests considering a more aggressive low-density lipoprotein cholesterol treatment goal in this group of individuals. In addition, the presence of the metabolic syndrome is highly predictive of the development of diabetes mellitus. A treatment strategy focusing on aerobic exercise and weight loss can help delay or prevent the development of diabetes and can help reduce cardiovascular risk. For significant risk reduction to be achieved, treatment strategies must focus on therapy for all risk factors, including dyslipidemia, hypertension, and insulin resistance.


An overview of coronaviruses including the SARS-2 coronavirus - Molecular biology, epidemiology and clinical implications.
Coronavirus infections have emerged as epidemic and pandemic threats in last two decades. After the H1N1 influenza pandemic in 2009, recently diagnosed novel betacoronavirus or severe acute respiratory syndrome coronavirus (SARS-CoV)-2 has spread across 203 countries and territories in all 5 major continents. World Health Organization (WHO) declared this as a public health emergency of international concern on January 30, 2020. Subsequently on February 11, 2020 a new name was given to this disease i.e. COVID-19 by an expert group from WHO. As of April 12, 2020, 10:00 CET, GMT+2:00, 1,696,588 confirmed cases and 105,952 confirmed deaths have been reported to the WHO. (Coronavirus disease 2019, situation report 83). It possibly originated from a small animal market in Wuhan, China. A cluster of patients were admitted with unusual pneumonia not responding to treatment in various hospitals. Epidemiological, genomic analysis and correlation with other coronaviruses led to the isolation of new coronavirus, closely resembling the bat coronaviruses, from such patients in Wuhan. They were identified as the SARS-CoV-2. This virus infection presents as influenza like illness in the affected people. Fever, cough, respiratory distress with fatigue, diarrhea, nausea and vomiting are common symptoms seen in adults. This may progress on to severe respiratory distress, hypoxia, need for oxygen supplementation and ventilator support as seen in patients in the SARS-CoV-1 epidemic (2003) in Guangdong, China. The transmissibility of SARS-CoV-1 was less as compared to SARS-CoV-2 infection, and it was well controlled with good public health efforts. The present COVID-19 epidemic is still in the acceleration phase of 3 and 4 in various countries. Without any effective antiviral agents available at present, the need of the hour is early case detection, isolation of cases, use of good preventive care measures by the household contacts and in the hospital set up. The results of ongoing clinical trials on hydroxychloroquine, azithromycin alone or in combination and a new antiviral agent remdesivir may help to treat some of the infections. A need for effective vaccine is being seen an as good preventive strategy in this pandemic. However the results of clinical trials and incorporation of vaccines in public health programs is a long way to go.


Ligand-induced interconversion of a coordination-organized porphyrin dimer: a potential fluorescence-based molecular memory monitor.
A novel interconversion system between less-fluorescent stacked (S) dimer and fluorescent extended (E) dimer of the monoimidazolylbisporphyrinatozinc complex was investigated. The addition of pyridine induces transformation from the S to the E dimer, whereas the addition of acetic acid and subsequent heating reverses the transformation. The interconversion rate is controlled by ligand concentration and thermal treatment. The system can be applied to repeatedly readable molecular memory by highly sensitive fluorescence detection.


Chronic traumatic encephalopathy: a potential late effect of sport-related concussive and subconcussive head trauma.
Chronic traumatic encephalopathy (CTE) is a form of neurodegeneration believed to result from repeated head injuries. Originally termed dementia pugilistica because of its association with boxing, the neuropathology of CTE was first described by Corsellis in 1973 in a case series of 15 retired boxers. CTE has recently been found to occur after other causes of repeated head trauma, suggesting that any repeated blows to the head, such as those that occur in American football, hockey, soccer, professional wrestling, and physical abuse, can also lead to neurodegenerative changes. These changes often include cerebral atrophy, cavum septi pellucidi with fenestrations, shrinkage of the mammillary bodies, dense tau immunoreactive inclusions (neurofibrillary tangles, glial tangles, and neuropil neurites), and, in some cases, a TDP-43 proteinopathy. In association with these pathologic changes, disordered memory and executive functioning, behavioral and personality disturbances (eg, apathy, depression, irritability, impulsiveness, suicidality), parkinsonism, and, occasionally, motor neuron disease are seen in affected individuals. No formal clinical or pathologic diagnostic criteria for CTE currently exist, but the distinctive neuropathologic profile of the disorder lends promise for future research into its prevention, diagnosis, and treatment.


Quantile Score: A New Reference System for Quantitative Fetal Echocardiography Based on a Large Multicenter Study.
Normative ranges of fetal echocardiographic measurements are important for quantitative diagnosis of fetal cardiovascular disease. The current normative ranges were derived from small samples and were based on the hypothesis of a normal distribution of these measurements during fetal cardiovascular growth. The aims of this study were to test the hypothesis of a normal distribution of fetal echocardiographic measurements in a large multicenter cohort and to propose a reference system without the normal distribution hypothesis to improve accuracy of fetal echocardiographic measurements. Fifty-two variables from 6,343 normal fetal echocardiographic examinations were acquired from seven Chinese centers. The hypothesis of a normal distribution used in ordinary least squares regression was tested with the Jarque-Bera test. The quantile score (q score) derived from quantile regression without normal distribution hypothesis was compared with the Z score derived from ordinary least squares regression. A total of 288 fetuses with outflow tract and great artery abnormalities and 300 normal fetuses were used to compare the diagnostic accuracy of q and Z scores. All fetal echocardiographic measurements showed non-normal distributions (P < .001). The normal range was underestimated by ordinary least squares regression compared with quantile regression by 30 ± 11%. The partial normalized areas under the receiver operating characteristic curve within the 20% false-positive rate were 0.62 and 0.50 for the q and Z scores, respectively. The q score provides a more robust system for determining normative ranges of fetal echocardiographic measurements. The improved sensitivity of matched false-positive rates makes the q score a more accurate reference for prenatal diagnosis, assessment, and prognosis of fetal cardiovascular disease.


The effect of intravenous dextrose infusion on postbypass hyperglycemia in pediatric patients undergoing cardiac operations.
To determine whether elimination of intraoperative dextrose-containing infusions affects post-cardiopulmonary bypass hyperglycemia in pediatric patients. Randomized, unblinded, saline-controlled study of perioperative glucose infusions in children undergoing cardiac surgery. Cardiac surgery suite and pediatric intensive care unit (ICU) of a university medical center. 33 consecutive, nondiabetic children undergoing cardiac surgery with deep hypothermia over an 8-month period. Group A (n = 16) received only normal saline infusions intraoperatively, and Group B (n = 17) received 5% dextrose and lactated Ringer's solution exclusively. Blood glucose was sampled immediately after induction of anesthesia, prior to cardiopulmonary bypass (CPB), after separation from CPB, on arrival in the ICU, and the morning of the first postoperative day. Data were analyzed using Student's t-test for independent samples, paired t-test, and analysis of variance, with p < 0.05 considered significant. Although moderate elevations in blood glucose (mean less than 165 mg/dl) after CPB were present in Group A, significantly higher levels (mean greater than 250 mg/dl) were noted in Group B. No children were hypoglycemic (glucose less than 40 mg/dl). Glucose levels were normal in both groups on the morning of the first postoperative day despite patients' having received continuous dextrose infusions in the ICU and the presumed stress of emergence from anesthesia. Extreme postbypass hyperglycemia can be controlled by eliminating intraoperative dextrose infusions. Hypoglycemia, an unlikely event, can easily be avoided by regular blood sampling, which is facilitated by the routine placement of arterial catheters.


Effect of extended trading hours on alcohol sales in Norway: study protocol for a stepped wedge cluster-randomized trial.
Norwegian alcohol policy measures include national restrictions on sales hours and a state monopoly on retail sales. A 1-hour extension of sales hours on Saturdays in the monopoly outlets took effect from September 2020. We aim to evaluate whether increase in sales hours results in (1) an increase in alcohol sales in the monopoly outlets and (2) an increase in total alcohol sales, including substitution effects from beer sales in grocery stores. The extension of Saturday sales hours is implemented within a stepped wedge cluster-randomized trial design. Block randomization of 62 of the 66 Norwegian trade districts allocated monopoly outlets to one of three sequences regarding date of implementation. A total of 228 of 335 in total Norwegian state monopoly outlets are eligible and included. The extension of sales hours is from 3 p.m. to 4 p.m. starting on the first Saturday in (i) September 2020, (ii) December 2020 or (iii) March 2021. Growth rates in monthly alcohol sales, measured in litres of pure alcohol, in eligible monopoly outlets (primary outcome) are obtained together with beverage-specific sales and alcohol sales in grocery stores (secondary outcomes). The observation period is set to 72 months prior to and 24 months after implementation. Power analyses indicate that this stepped wedge cluster-randomized controlled trial has a power above 90%, even at a high significance level (α = 0.01) and with other conservative model specifications. The planned trial offers a rare opportunity to study possible causal effects of a relatively small change in a widely used alcohol policy measure.


Clinical severe hepatic venoocclusive disease during induction treatment of acute monoblastic leukemia managed with defibrotide.
Hepatic venoocclusive disease (VOD) is the most common complication of cytoreductive therapy used for stem cell transplantation, but it is rarely encountered during induction treatment of acute leukemia. We describe a patient in whom severe clinical VOD developed shortly after induction treatment for acute monoblastic leukemia. Administration of intravenous defibrotide for 19 days induced complete resolution of the VOD. Further consolidation treatment was resumed including high-dose cytosine arabinoside without further complications.


Third case of Duchenne muscular dystrophy and West syndrome: Expanding the spectrum of the DMD neuropsychiatric phenotype.
Duchenne muscular dystrophy is an X-Linked neuromuscular disorder, and the most common muscular dystrophy. Neuropsychiatric phenotype associated to DMD gene mutations include now low IQ scores, epilepsy, autism, and attention deficit disorder. These have been observed with higher frequency in mutations that disrupt the short isoforms Dp71 and Dp140. West syndrome has been previously reported in two unrelated patients with Duchenne muscular dystrophy. Here, we report the third patient with West syndrome who had a novel hemizygous nonsense pathogenic variant in the exon 8 of the DMD gene c.811C>T, p.(Gln271*), suggesting West syndrome as part of the neuropsychiatric spectrum in Duchenne muscular dystrophy.


Isolation and culture of rhesus adipose-derived stem cells.
Adipose tissue is as an abundant and accessible source of stem cells with multipotent properties suitable for tissue engineering and regenerative medical applications. Rhesus monkeys are physiologically and phylogenetically similar to humans, and they and their cells are valuable for biomedical research and evaluation of preclinical therapies. Here, we describe methods for the isolation, culture, and differentiation of rhesus adipose-derived stem cells (rASCs).


4-{2-[(3,4-Dichloro-phen-yl)(meth-yl)amino]-4-methyl-1,3-thia-zol-5-yl}-N-(3-methyl-phen-yl)pyrimidin-2-amine.
In the title compound, C(22)H(19)Cl(2)N(5)S, the thia-zole and pyrimidine rings are almost co-planar, making a dihedral angle of 6.48 (7)°. In the crystal, intermolecular N-H⋯N hydrogen bonds link pairs of molecules into centrosymmetric dimers..


Post-operative Morbidity and Mortality After Fontan Procedure in Patients with Heterotaxy and Other Situs Anomalies.
Heterotaxy is a complex, multisystem disorder associated with single ventricle heart disease and decreased survival. Ciliary dysfunction is common in heterotaxy and other situs abnormalities (H/SA) and may increase post-operative complications. We hypothesized that patients with H/SA have increased respiratory and renal morbidities and increased in-hospital mortality after Fontan procedure. We queried the Pediatric Health Information System database for hospitalizations with ICD-9/10 codes for Fontan procedure in patients aged 1 through 11 years from 2004 to 2019. H/SA was identified by codes for dextrocardia, situs inversus, asplenia/polysplenia, or atrial isomerism and compared to non-H/SA controls. Outcomes were in-hospital mortality or heart transplantation, ECMO, hemodialysis, length of stay (LOS), and mechanical ventilation or vasoactive medication use ≥ 4 days. We adjusted estimates with multivariable logistic regression. Of 7897 patients at 50 centers, 1366 (17%) met criteria for H/SA. H/SA had worse outcomes for all study measures: death/transplantation (1.9 vs 1.1%, OR 1.74 (95% CI 1.01-3.03); p = 0.047), ECMO (3.7 vs 2.3%, OR 1.74 (1.28-2.35); p < 0.001), hemodialysis (2.1 vs 1.2%, OR 1.66 (1.06-2.59); p = 0.026), prolonged mechanical ventilation (13.2% vs 7.6%, OR 1.85 (1.53-2.25); p < 0.001) and vasoactive medication use (29.4 vs 19.7%, OR 1.65 (1.43-1.90), and longer LOS (11 (8-17) vs 9 (7-14) days; p < 0.001). H/SA is associated with increased cardiovascular, renal, and respiratory morbidity, as well as in-hospital mortality after Fontan procedure. Attention to renal and respiratory needs may improve outcomes in this difficult population. The relationship between ciliary dysfunction and lung and renal morbidity should be explored further.


Expression Profiling of Four Mango FT/TFL1-Encoding Genes under Different Fruit Load Conditions, and Their Involvement in Flowering Regulation.
Plant flowering is antagonistically modulated by similar FLOWERING LOCUS T (FT) and TERMINAL FLOWER 1 (TFL1) proteins. In mango (Mangifera indica L.), flowering is induced by cold temperatures, unless the tree is juvenile or the adult tree had a high fruit load (HFL) in the summer. Here, we studied the effects of juvenility and fruit load on the expression of four MiFT/TFL1 genes cloned from the mango 'Shelly' cultivar. Ectopic expression of MiFT1 in Arabidopsis resulted in early flowering, whereas over-expression of MiFT2 and the two cloned MiTFL1 genes repressed flowering. Moreover, juvenility was positively correlated with higher transcript levels of MiFT2 and both MiTFL1s. In trees with a low fruit load, leaf MiFT1 expression increased in winter, whereas HFL delayed its upregulation. MiFT2 expression was upregulated in both leaves and buds under both fruit load conditions. Downregulation of both MITFL1s in buds was associated with a decrease in regional temperatures under both conditions; nevertheless, HFL delayed the decrease in their accumulation. Our results suggest that cold temperature has opposite effects on the expression of MiFT1 and the MiTFL1s, thereby inducing flowering, whereas HFL represses flowering by both suppressing MiFT1 upregulation and delaying MiTFL1s downregulation. The apparent flowering-inhibitory functions of MiFT2 are discussed.


High Throughput Synthesis of 2,3,6-Trisubstituted-5,6-Dihydroimidazo[2,1-b]thiazole Derivatives.
A facile approach to the synthesis of 2,3,6-trisubstituted-5,6-dihydroimidazo[2,1-b]thiazole was reported. A resin bound cyclic thiourea was formed by the treatment of a resin bound diamine with 1,1'-thiocarbonyldiimidazole, and then reacted with a α-haloketone to generate a resin bound isothiourea. HF treatment of the resin bound isothiourea resulted in the cleavage of the product and simultaneous formation of an enamine bond. This led to the formation of the 2,3,6-trisubstituted-5,6-dihydroimidazo[2,1-b]thiazole in high yield and purity.


Cortical Circuit Dynamics Are Homeostatically Tuned to Criticality In Vivo.
Homeostatic mechanisms stabilize neuronal activity in vivo, but whether this process gives rise to balanced network dynamics is unknown. Here, we continuously monitored the statistics of network spiking in visual cortical circuits in freely behaving rats for 9 days. Under control conditions in light and dark, networks were robustly organized around criticality, a regime that maximizes information capacity and transmission. When input was perturbed by visual deprivation, network criticality was severely disrupted and subsequently restored to criticality over 48 h. Unexpectedly, the recovery of excitatory dynamics preceded homeostatic plasticity of firing rates by >30 h. We utilized model investigations to manipulate firing rate homeostasis in a cell-type-specific manner at the onset of visual deprivation. Our results suggest that criticality in excitatory networks is established by inhibitory plasticity and architecture. These data establish that criticality is consistent with a homeostatic set point for visual cortical dynamics and suggest a key role for homeostatic regulation of inhibition.


Third-party verification of total and specific immunoglobulin E on analyzer Immulite® 2000XPi.
Although Immulite®2000XPi (Siemens) allergy testing is routinely used in laboratories worldwide; a well-designed comprehensive third-party verification of its analytical performance has not yet been published. Our aim was to verify stability, precision and reference intervals for total and some frequent specific IgEs, test trueness for several most common allergens and assess the comparability of fluorescent- and chemiluminescent-enzyme immunoassay for total IgE. Verification was based on EP15-A3, EP9-A3 and EP28-A3c CLSI guidelines. Stability was tested on serum pools stored at 2-8 °C, during 7 days. Manufacturer's reference intervals were verified for children and adults. Acceptance criteria were based on manufacturer specifications and national External Quality Assessment provider data. Samples for total IgE analysis were stable at 2-8 °C for 7 days. Precision was within manufacturer's specifications for most of tested allergens, except for total IgE (92.5 kIU/L, CVs 5.6% and 7.6%) and w1 (9.3 kU/L, CV = 5.9%). Bland-Altman plot revealed statistically significant constant (-32.4 kIU/L) and proportional bias (-20.2%) for total IgE between two methods. Passing Bablok showed statistically but not clinically significant biases between two Immulite analyzers for betula verrucosa (constant and proportional), house dust mite and egg white allergens (proportional). For adults and children, total IgE results were within reference intervals declared by Siemens. Immulite®2000XPi assay has acceptable precision and trueness. Samples for total IgE are stable even longer than declared by the manufacturer. Declared reference intervals can safely be adopted for both adult and children population in our laboratory. Fluorescent- and chemiluminescent-enzyme immunoassay results of total IgE are not comparable.


Optimized serum stability and specificity of an αvβ6 integrin-binding peptide for tumor targeting.
The integrin αvβ6 is an antigen expressed at low levels in healthy tissue but upregulated during tumorigenesis, which makes it a promising target for cancer imaging and therapy. A20FMDV2 is a 20-mer peptide derived from the foot-and-mouth disease virus that exhibits nanomolar and selective affinity for αvβ6 versus other integrins. Despite this selectivity, A20FMDV2 has had limited success in imaging and treating αvβ6+ tumors in vivo because of its poor serum stability. Here, we explore the cyclization and modification of the A20FMDV2 peptide to improve its serum stability without sacrificing its affinity and specificity for αvβ6. Using cysteine amino acid substitutions and cyclization by perfluoroarylation with decafluorobiphenyl, we synthesized six cyclized A20FMDV2 variants and discovered that two retained binding to αvβ6 with modestly improved serum stability. Further d-amino acid substitutions and C-terminal sequence optimization outside the cyclized region greatly prolonged peptide serum stability without reducing binding affinity. While the cyclized A20FMDV2 variants exhibited increased nonspecific integrin binding compared with the original peptide, additional modifications with the non-natural amino acids citrulline, hydroxyproline, and d-alanine were found to restore binding specificity, with some modifications leading to greater αvβ6 integrin selectivity than the original A20FMDV2 peptide. The peptide modifications detailed herein greatly improve the potential of utilizing A20FMDV2 to target αvβ6 in vivo, expanding opportunities for cancer targeting and therapy.


Multimodal cues displayed by submissive rats promote prosocial choices by dominants.
Animals often display prosocial behaviors, performing actions that benefit others. Although prosociality is essential for social bonding and cooperation, we still know little about how animals integrate behavioral cues from those in need to make decisions that increase their well-being. To address this question, we used a two-choice task where rats can provide rewards to a conspecific in the absence of self-benefit and investigated which conditions promote prosociality by manipulating the social context of the interacting animals. Although sex or degree of familiarity did not affect prosocial choices in rats, social hierarchy revealed to be a potent modulator, with dominant decision-makers showing faster emergence and higher levels of prosocial choices toward their submissive cage mates. Leveraging quantitative analysis of multimodal social dynamics prior to choice, we identified that pairs with dominant decision-makers exhibited more proximal interactions. Interestingly, these closer interactions were driven by submissive animals that modulated their position and movement following their dominants and whose 50-kHz vocalization rate correlated with dominants' prosociality. Moreover, Granger causality revealed stronger bidirectional influences in pairs with dominant focals and submissive recipients, indicating increased behavioral coordination. Finally, multivariate analysis highlighted body language as the main information dominants use on a trial-by-trial basis to learn that their actions have effects on others. Our results provide a refined understanding of the behavioral dynamics that rats use for action-selection upon perception of socially relevant cues and navigate social decision-making.


Transforming growth factor-β receptor 1: An intervention target for genetic poor cartilage quality induced by prenatal dexamethasone exposure.
Fetal-originated osteoarthritis is relative to poor cartilage quality and may exhibit transgenerational genetic effects. Previous findings revealed prenatal dexamethasone exposure (PDE) induced poor cartilage quality in offspring. This study focused on further exploring molecular mechanism, heritability, and early intervention of fetal-originated osteoarthritis. Pregnant rats (F0) were segregated into control and PDE groups depending upon whether dexamethasone was administered on gestational days (GDs) 9-20. Some female offspring were bred with healthy males during postnatal week (PW) 8 to attain the F2 and F3 generations. The F3-generation rats were administrated with glucosamine intragastrically at PW12 for 6 weeks. The knee cartilages of male and female rats at different time points were harvested to assay their morphologies and functions. Furthermore, primary chondrocytes from the F3-generation rats were isolated to confirm the mechanism and intervention target of glucosamine. Compared with the control, female and male rats in each generation of PDE group showed thinner cartilage thicknesses; shallower and uneven staining; fewer chondrocytes; higher Osteoarthritis Research Society International scores; and lower mRNA and protein expression of SP1, TGFβR1, Smad2, SOX9, ACAN and COL2A1. After F3-generation rats were treated with glucosamine, all of the above changes could be reversed. In primary chondrocytes isolated from the F3-generation rats of PDE group, glucosamine promoted SP1 expression and binding to TGFβR1 promoter to increase the expression of TGFβR1, p-Smad2, SOX9, ACAN and COL2A1, but these were prevented by SB431542 (a potent and selective inhibitor of TGFβR1). PDE induced chondrodysplasia in offspring and stably inherited in F3-generation rats, which was related to decreased expression of SP1/TGFβR1/Smad2/SOX9 pathway to reduce the cartilage matrix synthesis, without major sex-based variations. Glucosamine could alleviate the poor genetic cartilage quality in offspring induced by PDE by up-regulating SP1/TGFβR1 signaling, which was prevented by a TGFβR1 inhibitor. This study elucidated the molecular mechanism and therapeutic target (TGFβR1) of genetic chondrodysplasia caused by PDE, which provides a research basis for precisely treating fetal-originated osteoarthritis.


HCA587 Protein Vaccine Induces Specific Antitumor Immunity Mediated by CD4+ T-cells Expressing Granzyme B in a Mouse Model of Melanoma.
The antigen HCA587 (also known as MAGE-C2), which is considered a cancer-testis antigen, exhibits upregulated expression in a wide range of malignant tumors with unique immunological properties, and may thus serve as a promising target for tumor immunotherapy. The study aimed to explore the antitumor effect of the HCA587 protein vaccine and the response of humoral and cell-mediated immunity. The HCA587 protein vaccine was formulated with adjuvants CpG and ISCOM. B16 melanoma cells were subcutaneously inoculated to C57BL/6 mice, followed by treatment with HCA587 protein vaccine subcutaneously. Mouse survival was monitored daily, and tumor volume was measured every 2 to 3 days. The tumor sizes, survival time and immune cells in tumor tissues were detected. And the vital immune cell subset and effector molecules were explored. After treatment with HCA587 protein vaccine, the vaccination elicited significant immune responses, which delayed tumor growth and improved animal survival. The vaccination increased the proportion of CD4+ T cells expressing IFN-γ and granzyme B in tumor tissues. The depletion of CD4+T cells resulted in an almost complete abrogation of the antitumor effect of the vaccination, suggesting that the antitumor efficacy was mediated by CD4+ T cells. In addition, knockout of IFN-γ resulted in a decrease in granzyme B levels, which were secreted by CD4+ T cells, and the antitumor effect was also significantly attenuated. The HCA587 protein vaccine may increase the levels of granzyme B expressed by CD4+ T cells, and this increase is dependent on IFN-γ, and the vaccine resulted in a specific tumor immune response and subsequent eradication of the tumor.


Differentiation capacities of the labial imaginal disc ofDrosophila melanogaster.
The mature labial disc, when implanted into a larva of the same age, undergoes metamorphosis along with the host and produces one lateral half of the medi- and distiproboscis. On the basis of results obtained from transplanted disc halves (including the separate peripodial membrane) a tentative fate map of the labial disc was constructed, which shows most of the presumptive mediproboscis to be located in the dorsal, and most of the presumptive distiproboscis in the ventral part of the disc. The distal protion of the peripodial membrane also contains imaginal anlagen, viz. part of the mediproboscis, prementum, and labellar cap anlagen. The involvement of this part of the peripodial membrane was checked by a careful histological analysis of labial disc development during the first ten hours after prepupation. The results were compared with the situation described forCalliphora imaginal discs.In addition, a detailed morphological analysis was made of the proboscis of the homoeotic mutantproboscipedia (pb). At 27°C,pb changes the distiproboscis into a "telopodite" (leg segments distal to the coxa); the (unchanged) prementum may therefore correspond to the coxa. At 15° C, the tarsus of this homoeotic "telopodite" is replaced to a greater or lesser extent by an arista. The present analysis thus confirms (a) the fundamental morphological correspondence of the medi- and distiproboscis with the labium of other insects, and (b) the fundamental developmental correspondence of the labial, antennal, and leg discs.


Transient expression of a 'lens-specific' gene, delta-crystallin, in the embryonic chicken adenohypophysis.
A characteristic protein of the lens, delta-crystallin, has been reported previously to be present in the embryonic chicken adenohypophysis. We confirmed this earlier finding by biochemical detection of delta-crystallin protein using a monoclonal antibody and delta-crystallin mRNA using a specific cDNA probe. We estimate the concentration of delta-crystallin and its mRNA in the 3.5-day embryonic chicken adenohypophysis to be approximately 1/3,000 and 1/5,000 of the respective value found in lens. Tissue culture revealed that cells positive for delta-crystallin comprise about 30% of embryonic adenohypophysis and are randomly scattered in this organ. No lentoid formation was observed during the culture period.


Stakeholder Power Analysis of the Facilitators and Barriers for Telehealth Solution Implementation in China: A Qualitative Study of Individual Users in Beijing and Interviews With Institutional Stakeholders.
Facing COVID 19, the use of telehealth solutions grows exponentially. However, despite the large investments made into telehealth solutions, the implementation process remains slow and sluggish. Moreover, during COVID-19, older people experienced difficulties and had the highest mortality rates, and those lucky enough to survive faced tremendous pressure to use QR code-based health monitoring systems. This paper aims to determine the barriers and incentives for the implementation of telehealth solutions via a case study about telehealth implementation in China. We conducted 8 semi-structured interviews following the design of the interactive learning framework (research question defining, participant recruitment, exploratory stage, consultation stage, integration stage, and follow-up interview). One interview with a government official from the National Health Commission and another interview with a government official from the China Center for Disease Control and Prevention was conducted in the exploratory stage. The consultation stage comprised one interview with a business manager from the Huawei Wearable Unit, one interview with a business manager from Alibaba Health Brain Unit, and one interview with a business manager from Xiaomi. Two interviews with doctors from Fudan University-affiliated Huashan Hospital and Fudan University-affiliated Zhongshan hospital were conducted in the integration stage. In addition, 8 focus group studies with 64 participants from rural and urban Beijing were conducted. Finally, another telephone interview with a business manager of the Xiaomi Wearable Unit was conducted in the follow-up stage. Telehealth solutions are designed to assist health care providers in realizing the quadruple aim of better health outcomes, lowering health care costs, improved health care quality, and improved doctor and patient experiences. Governments have high incentives to improve health care efficiency via telehealth solutions. However, they have limited resources to make the necessary infrastructure transformation. To fully realize the potential of telehealth devices, heavy infrastructure investment in the telecommunication network is required beforehand to resolve the interoperability issue occurring during the data collection process for telehealth solutions. The industry also demands a mature business model incorporating collaboration between various stakeholders and industrial partners to invest in infrastructure. Governments have high interest and significant influence on building the necessary infrastructure for telehealth solution implementation in China. Industrial actors have a high interest and a medium level of power for telehealth solution implementation. Users have high interest but a lower level of power for the usage of telehealth solutions, and doctors have low interest and a medium level of power for telehealth solutions implementation.


Intramolecular dielectric screening in proteins.
This paper investigates the microscopic mechanisms of charge screening by proteins. For this purpose, we introduce the generalized susceptibility of a protein in response to a point charge, which is a scalar quantity dependent on position within the protein. The contribution to the susceptibility from atomic polarizabilities, associated with electronic degrees of freedom, is found to be highly uniform. By contrast, that from dynamic dipolar relaxation, associated with nuclear degrees of freedom, varies greatly between different regions of the protein. We investigate the possible rôle of this variation in the activity of proteins that interact functionally with charged species, and we formulate and test the hypothesis that this variation is correlated to functional activity. Model calculations give encouraging support to this hypothesis. The protein's dielectric properties are represented by a standard model in which electronic relaxation is described by a set of atomic polarizabilities, and dipolar relaxation is treated as a perturbation to normal mode dynamics. The model yields the desired susceptibility in closed form. Its obvious limitations are discussed. It is applied to several test systems, and is compared to various continuum models. Four model alpha-helices are considered, three of which play a rôle in vivo in the binding of charged ligands. We show that the intramolecular screening, and its spatial variation, can indeed play a part in this binding. The electron transfer between ferri- and ferrocytochrome c is considered. The dielectric relaxation of each molecule, associated respectively with its oxidation or its reduction, is known to be directly related to the activation free energy for the electron transfer reaction. Our analysis of the dielectric susceptibility will thus permit an estimate of this activation free energy. We show that the relaxation of the atomic positions ("dipolar relaxation") contributes 1 kcal/mol to this activation free energy, and that the molecule achieves this low value by providing a low dipolar susceptibility throughout its central part. In this case, the spatial variation of the susceptibility has a clear functional rôle.