Metabolomic and transcriptomic changes underlying cold and anaerobic stresses after storage of table grapes.
The currently accepted paradigm is that fruits and vegetables should be consumed fresh and that their quality deteriorates during storage; however, there are indications that some metabolic properties can, in fact, be improved. We examined the effects of low temperature and high-CO2 conditions on table grapes, Vitis vinifera L. cv. 'Superior Seedless'. Berries were sampled at harvest (T0) and after low-temperature storage for 6 weeks under either normal atmosphere conditions (TC) or under an O2 level of 5 kPa and elevated CO2 levels of 5, 10 or 15 kPa (T5, T10, T15). Accumulation of 10 stilbenes, including E-ε-viniferin, E-miyabenol C and piceatannol, significantly increased under TC treatment as compared to T0 or T15. Sensory analysis demonstrated that elevated CO2 elicited dose-dependent off-flavor accumulation. These changes were accompanied by an accumulation of 12 volatile metabolites, e.g., ethyl acetate and diacetyl, that imparted disagreeable flavors to fresh fruit. Transcriptome analysis revealed enrichment of genes involved in pyruvate metabolism and the phenylpropanoid pathway. One of the transcription factors induced at low temperature but not under high CO2 was VvMYB14, which regulates stilbene biosynthesis. Our findings reveal the potential to alter the levels of targeted metabolites in stored produce through understanding the effects of postharvest treatments.


Long-term cognitive outcomes in term newborns with watershed injury caused by neonatal encephalopathy.
We previously reported that increasing severity of watershed (WS) injury in neonatal magnetic resonance imaging (MRI) is associated with worse language outcomes in early childhood. In the present study, we investigated the relationship between neonatal injury patterns and cognitive profile in adolescents with neonatal encephalopathy. Term neonates with encephalopathy were prospectively enrolled and imaged using brain MRI from 1999 to 2008. Neonatal brain injury was scored according to the degree of injury in WS and basal ganglia/thalamus (BG/T) areas. The children underwent a neurocognitive assessment and follow-up brain MRI at the age of 10-16 years. The relationship between neonatal brain injury patterns and adolescent cognitive outcomes was assessed. In a cohort of 16 children, neonatal MRI showed WS injury in 7, BG/T injury in 2, and normal imaging in 7. Children with WS injury had lower estimated overall cognitive ability than those with normal imaging. Increasing WS injury score was associated with decreasing estimated overall cognitive ability, Perceptual Reasoning Index, and digit span score. Children with the WS injury are at an increased risk of having problems in long-term intellectual ability. These cognitive outcomes may underlie early language difficulties seen in children with neonatal WS injury. Adolescents with a history of neonatal encephalopathy and watershed pattern of injury on neonatal brain magnetic resonance imaging (MRI) had lower overall cognitive ability, perceptual reasoning skills, and auditory working memory than those with normal neonatal imaging. Children with post-neonatal epilepsy and cerebral palsy had the worst cognitive outcomes. Watershed pattern of injury confers high long-term differences in intellectual ability.


Global landscape of lysine acylomes in Bacillus subtilis.
Lysine acetylation is a common posttranslational modification that regulates numerous biochemical functions in both eukaryotic and prokaryotic species. In addition, several new non-acetyl acylations are structurally different from lysine acetylation and participate in diverse physiological functions. Here, a comprehensive analysis of several lysine acylomes was performed by combining the high-affinity antibody enrichment with high-resolution LC-MS/MS. In total, we identified 2536 lysine acetylated sites, 4723 propionylated sites, 2150 succinylated sites and 3001 malonylated sites in Bacillus subtilis, respectively. These acylated proteins account for 35.8% of total protein in this bacterium. The four lysine acylomes showed a motif preference for glutamate surrounding the modified lysine residues, and a functional preference for several metabolic pathways, such as carbon metabolism, fatty acid metabolism, and ribosome. In addition, more protein-protein interaction clusters were identified in the propionylated substrates than other three lysine acylomes. In summary, our study presents a global landscape of acylation in the Gram-positive model organism Bacillus and their potential functions in metabolism and physiology.


Physico-chemical characterization and pharmacological activities of polysaccharides from Opuntia microdasys var. rufida cladodes.
The aim of this study is to isolate pectin from peel (WNPE) and pulp (WNPU) of Opuntia microdasys var. rufida's (OMR) cladodes and to characterize these polysaccharides by size exclusion (SEC/MALS/VD/DRI), gas chromatography coupled to mass spectrometer (GC-MS), nuclear magnetic resonance (1H NMR), and Fourier transform infrared spectroscopy (FTIR) analysis. The polysaccharides were extracted in neutral aqueous media followed by ethanol precipitation and dialysis. Both WNPE and WNPU are mainly composed of uronic acids and some neutral sugars such as arabinose, galactose, rhamnose and mannose. Their molecular weight range from 2,180,000 and 4,920,000g/mol. The in-vivo pharmacological activities (anti-inflammatory, analgesic and gastroprotective activities) have been performed. The extracted pectin (50-100mg/kg, i.p. (intraperitoneal)) inhibited, in a dose-related manner, both carrageenan-induced paw edema in rats and Xylene-induced ear edema in mice. A dose-dependent action was obtained against chemical (writhing test) and thermic (hot plate test) stimuli, respectively, with doses of 50 and 100mg/kg. Moreover, a considerable gastroprotective effect was observed with these two biopolymers, the gastric ulcer was attenuated until 67.67% for WNPE and 81.93% for WNPU, at the dose of 100mg/kg.


Identification of transfer RNA suppressors in Escherichia coli. III. Ochre suppressors of lysine tRNA.
Transducing phages of lambda carrying suppressors, lysT (Su+ beta), supG and and supL, were isolated in vivo. Upon infection with each of these phages, the production of tRNALys and tRNAVal1 was markedly enhanced. Fingerprint analysis of these tRNAs revealed that they consisted of normal tRNALys, mutant tRNALys and tRNAVal1 in equimolar ratios. The mutant tRNALys carried a single-base alteration at the anticodon, from 5'-UUU-3' to 5'-UUA-3', which makes it an ochre suppressor. DNA sequence analysis of the entire transducing fragment (730 base-pairs) of lambda pSu+ beta revealed that three tRNA genes are tightly clustered within a transcription unit in the following order; i.e. promoter-(48 base-pairs)-wild-type tRNALys-(132 base-pairs)-tRNAVal1-(2 base-pairs)-Su+ beta tRNALys-. In wild-type bacteria there are two identical tRNALys genes in one operon. Although we have shown that in Su+ beta it is the distal tRNALys that has been mutated to the ochre suppressor by a single base change at the anticodon (U36 to A36), we have not determined which of the two genes bears the supG or the supL mutation. The sequences following both tRNALys genes are highly homologous: both are about 100 base-pairs long and both terminate with an 18 base-pair sequence homologous to the last 18 bases of each tRNA. The sequences of tRNALys and tRNAVal1 are also very similar. Thus, including the 3'-portions of these tRNA genes, the 18 base-pair sequence is more or less periodically repeated five times in the DNA sequence.


Prognostic factors for patients with chronic myeloid leukaemia in chronic phase treated with imatinib mesylate after failure of interferon alfa.
We assessed clinical results in 145 patients with chronic myeloid leukaemia in chronic phase who satisfied criteria for interferon-alpha failure and were thus eligible for treatment with imatinib at the Hammersmith Hospital. We used univariate and multivariate analyses to develop a risk score based on features defined after treatment for 3 months. We identified a low neutrophil count and poor cytogenetic response (<35% Ph-negative marrow metaphases) at 3 months as principal independent predictive factors and incorporated them into a three-tier prognostic scoring system for individual patients. For patients in the low-, intermediate- and high-risk groups, the probabilities of survival at 24 months were 100, 82 and 40% (P<0.0001) and progression-free survival 100, 66 and 15% (P<0.0001), respectively. This Hammersmith prognostic scoring system was validated with an independent cohort of patients treated at another UK centre.


Low temperature extrusion blown ε-polylysine hydrochloride-loaded starch/gelatin edible antimicrobial films.
Edible antimicrobial films made from starch/gelatin (S/G) incorporated with different ε-polylysine hydrochloride (ε-PL) contents were developed by low-temperature extrusion blowing process. ε-PL addition reduced the complex viscosity and storage modulus of blends, while promoted the formation of hydrogen bonding among film components. The control film had an A-type crystalline structure, while increasing the ε-PL content promoted its transformation to B-shaped structure. Without ε-PL and under the processing temperature used, the starch granules were not sufficiently gelatinized. However, ε-PL addition significantly enhanced the gelatinization degree. Increasing ε-PL content in S/G films increased film flexibility, water contact angle value, swelling degree, antimicrobial effect, and storage period of fresh bread, but decreased water vapor permeability and tensile strength. S/G film with 4 wt% ε-PL had the highest water contact angle (94°) and elongation at break (149%). This research demonstrates the plasticizing effects of ε-PL and potential of S/G films containing ε-PL for food preservation/packaging.


Advice for young investigators: historical perspectives on scientific research.
Physicians attempting to carry out well-executed scientific investigations require an historical perspective; an awareness of our ignorance; an imagination unencumbered by scientific prejudices; originality, curiosity and imagination; patience and persistence in the face of frustration; adaptability and perseverance; a critical spirit of evaluation and willingness to question dogma; devotion to the search for truth; and an awareness of those responsibilities to individuals and to society which define the medical profession. This paper explores, through the words of famous scientists, philosophers and authors, the process of scientific investigation and discusses those qualities that investigators can develop to become more insightful and successful researchers.


Two cases of intratemporal facial nerve neurilemmomas.
The patients with neurilemmomas of the facial nerve within the temporal bone were treated. Although most of cases of the facial nerve neurilemmoma in the temporal bone present with a facial palsy, a few cases have normal facial nerve function and are especially difficult to diagnose preoperatively because of few symptoms. Case 1, a 31-year-old man, developed progressive left facial palsy with protrusion of the posterior wall of the external auditory meatus over a five-month period. Case 2, a 64-year-old man, complained of only fullness and tinnitus of the right ear without any facial nerve symptoms. Polytomography of the temporal bone demonstrated no abnormal findings in both cases. However, computed tomography revealed a soft-tissue density area in the left mastoid cavity with destruction of the posterior wall of the external auditory meatus in case 1. A similar soft-tissue density mass was found in the right tympanic cavity in case 2, suggesting the presence of the tumor in the temporal bone. At operation, the tumors were found to originate from the vertical portion of the facial nerve in both cases. It was assumed that normally existing dehiscences of fallopian canal might account for why some patients have normal facial nerve function.


Walking Meeting Effects on Productivity and Mood Among White-Collar Workers: Evidence From the Walking Meeting Pilot Study.
Walking Meetings (WaM) have shown to be acceptable, feasible, and implementable among white-collar workers. This pilot study examines the effects of WaM on worker mood, productivity, and physical activity. Eighteen white-collar workers were recruited from a university in Florida. Participants wore accelerometers for 3 consecutive weeks. During baseline week, participants continued their normal seated meetings. In weeks 2 and 3, participants conducted one walking meeting in groups of two to three people. Increased moderate occupational physical activity was correlated with increased productivity measured by decreased work time missed due to health reasons (r = -0.59; P = 0.03) and decreased impairment while working due to health reasons (r = -0.61; P = 0.02). In week 3, very vigorous physical activity was significantly correlated with negative mood (r = 0.69, P = 0.01). WaM affected worker mood and productivity and increased occupational physical activity-levels.


Financial Hardship, Healthcare Utilization, and Health Among U.S. Cancer Survivors.
This study examined associations of both medical and nonmedical financial hardships with healthcare utilization and self-rated health among cancer survivors. The National Health Interview Survey (2013-2017) was used to identify cancer survivors (aged 18-64 years: n=4,939; aged ≥65 years: n=6,972). A total of 4 levels of medical financial hardship intensities were created with measures from material, psychological, and behavioral domains. A total of 5 levels of nonmedical financial hardship intensities were created with measures in food insecurity and worry about other economic needs (e.g., housing expenses). Generalized ordinal logistic regression examined associations between medical and nonmedical financial hardship intensities and emergency department visits, use of preventive services and cancer screenings, and self-rated health. All analyses were performed in 2019. In adjusted analyses, cancer survivors with higher medical financial hardship intensity (Level 4 vs Level 1; aged 18-64 years: 42% vs 26.2%, p<0.001; aged ≥65 years: 37.6% vs 24.3%, p=0.001) and higher nonmedical financial hardship intensity (Level 5 vs Level 1; aged 18-64 years: 37.2% vs 27.9%, p=0.011) had more emergency department visits. Moreover, cancer survivors with higher medical financial hardship intensity had lower influenza vaccine (Level 4 vs Level 1; aged 18-64 years: 45.6% vs 52.5%, p=0.036; aged ≥65 years: 64.6% vs 75.6%, p=0.008) and lower breast cancer screening levels (Level 4 vs Level 1; 46.8% vs 61.2%, p=0.001). Similar patterns were found between higher financial hardship intensities and worse self-rated health. Higher medical and nonmedical financial hardships are independently associated with more emergency department visits, lower receipt of some preventive services, and worse self-rated health in cancer survivors. With growing healthcare costs, unmet medical and nonmedical financial needs may worsen health disparities among cancer survivors.


Efficacy of a triamcinolone acetonide-loaded dissolving microneedle patch for the treatment of hypertrophic scars and keloids: a randomized, double-blinded, placebo-controlled split-scar study.
The treatment of hypertrophic scars (HTSs) and keloids remains a challenge. Intralesional triamcinolone acetonide (TAC) is the mainstay treatment for these conditions. Despite its efficacy, TAC has several adverse side effects, including telangiectasias, skin atrophy, pigmentary changes, and skin necrosis. Dissolving microneedles (DMN) use the poke-and-release method to create microchannels that enhance drug delivery to the target tissue in the dermis, without causing pain and with a decreased risk of transmission of blood-borne diseases. To evaluate and compare the efficacy of a TAC-DMN versus a drug-free DMN patch for the treatment of HTSs and keloids, 20 patients (10 with HTSs and 10 with keloids) received a split-scar treatment: one half of the scar length was treated with TAC-DMNs and the other half was treated with drug-free DMN for three sessions at 14-day intervals. Efficacy was assessed by measuring the scar volume through a multispectral imaging system and using the Patient and Observer Scar Assessment Scale (POSAS). The HTSs treated with TAC-DMNs showed a significant reduction in the mean scar volume 2 weeks after the second treatment and 1 month after the third treatment (p = 0.028 and 0.020, respectively), while the HTSs treated with drug-free DMNs showed no significant reduction in the scar volume. Both sides of the keloids showed no significant reduction in mean scar volume. Using the POSAS, significant improvement in the appearance of both halves of the HTSs was observed 1 month after the treatments. A significant improvement (evaluated by POSAS) was also observed in the keloids treated with TAC-DMNs 2 weeks after the second treatment and 1 month after the third treatment. No significant improvement was observed from the patients' perspective as evaluated by POSAS in the keloids treated with drug-free DMNs. However, no significant difference was observed between the treatment and control halves. TAC-DMN is an effective treatment for HTSs. Increasing the dosage and duration of keloid scar treatment is required in future studies to determine whether it would result in a significant therapeutic outcome. This trial is registered in the Thai Clinical Trials Registry (TCTR20220318004; date of registration, March 17, 2022).


Roadside Rhododendron pulchrum leaves as bioindicators of heavy metal pollution in traffic areas of Okayama, Japan.
The leaves of roadside Rhododendron pulchrum Sweet were examined as a bioindicator of traffic-related heavy metal pollution in Okayama. Total contents of heavy metals in roadside soil and the R. pulchrum leaves were determined. Results of correlation analyses showed significant correlations among Pb, Ni, and Zn contents of roadside soil and leaf samples. These results suggest that R. pulchrum is a useful bioindicator of heavy metal pollution in traffic areas. To identify traffic-related heavy metal pollution sources, traffic-related materials such as tires, asphalt pavement, and road paint were collected and analyzed. The results of analyses of our data show that Zn is emitted mainly from the abrasion of tires; Cr is emitted mainly from wear of the asphalt pavement. Furthermore, the respective lead isotope ratios in R. pulchrum leaves, soil, roadside dust, and traffic-related materials were measured using inductively coupled plasma mass spectrometry. The isotopic compositions of lead in R. pulchrum leaves were 207Pb/206Pb = 0.861-0.871 and 208Pb/206Pb = 2.093-2.127, which agrees well with ratios in automobile wheel balance weights, suggesting that lead-containing products associated with automobiles, such as wheel balance weights, contribute to traffic-related lead pollution.


Evaluation of GM1 ganglioside-mediated apoptosis in feline thymocytes.
Cats with inherited GM1 gangliosidosis (GM1 mutant cats) have premature thymic involution characterized by decreased total thymocytes primarily affecting the CD4+ CD8+ subpopulation. While GM1 mutant cats have increased cell surface GM1 gangliosides, as determined by cholera toxin B binding, on both thymocytes and peripheral lymph node cells only thymocytes show increased apoptosis. To determine if GM1 gangliosides can increase the occurrence of apoptosis in feline thymocytes directly, we added exogenous GM1 ganglioside (GM1) to feline thymocyte primary cultures and compared the results to apoptotic changes seen in untreated cells or in cells treated with dexamethasone (Dex), a known inducer of thymocyte apoptosis in other species. Incorporation of exogenous GM1 into thymocyte cytoplasmic membranes was confirmed by flow cytometric analyses of cholera toxin B labelling. Apoptosis in feline thymocytes was analyzed by electron microscopy, spectrophotometric evaluation of DNA fragmentation, flow cytometric enumeration of apoptotic nuclei, and gel electrophoretic analysis of degraded DNA. Alterations in percentages of thymocyte immunophenotype following GM1 incorporation were determined by flow cytometric analyses of labelled cell surface markers for feline CD4 and CD8. Because in vitro addition of GM1 gangliosides has been reported in other species to decrease surface expression of CD4 on both thymocytes and peripheral lymphocytes, we evaluated GM1-associated down-regulation of CD4 on the surface of feline thymocytes and peripheral lymph node cells by flow cytometry. Additionally, we compared the apoptotic response of the more mature peripheral lymph node cells to the less mature thymocytes. Our results indicate that incorporation of exogenous GM1 into feline thymocyte cell membranes produces a dose-dependent increase of apoptotic cell death. Although, CD4 expression on both feline thymocyte and lymph node cell membranes was abruptly decreased after introducing exogenous GM1, enhanced apoptotic death was observed only in thymocytes, not in lymph node cells at the same GM1 concentration. Enhancement of thymocyte apoptosis appears to be age-related since cells derived from cats <3 months of age were more vulnerable than those from cats >3 months of age.


Feedback regulation of ornithine decarboxylase expression. Studies using a polysomal run-off system.
The rate-controlling enzyme in polyamine synthesis, ornithine decarboxylase (ODC), is subject to feedback regulation by the polyamines at the level of translation. In the present study we used a cell-free translation system to further investigate the mechanism by which this regulation occurs. Lysates of ODC-overproducing cells were capable of synthesizing large amounts of ODC. The degree of initiation was poor in the lysates and the synthesis of ODC was mainly a result of continued elongation of peptide chains on pre-initiated ribosomes. By determining the amount of ODC produced in the lysate, we obtained an estimate of the number of ribosomes that were actively translating ODC mRNA at the moment of lysis. Using this polysomal run-off assay we demonstrated that the polyamine-mediated regulation of ODC synthesis occurs without any change in the number of ribosomes associated with the message. This finding indicates that the polyamines exert a coordinate effect on initiation and elongation.


A single fungal strain was the unexpected cause of a mass aspergillosis outbreak in the world's largest and only flightless parrot.
Kākāpō are a critically endangered species of parrots restricted to a few islands off the coast of New Zealand. Kākāpō are very closely monitored, especially during nesting seasons. In 2019, during a highly successful nesting season, an outbreak of aspergillosis affected 21 individuals and led to the deaths of 9, leaving a population of only 211 kākāpō. In monitoring this outbreak, cultures of aspergillus were grown, and genome sequenced. These sequences demonstrate that, very unusually for an aspergillus outbreak, a single strain of aspergillus caused the outbreak. This strain was found on two islands, but only one had an outbreak of aspergillosis; indicating that the strain was necessary, but not sufficient, to cause disease. Our analysis provides an understanding of the 2019 outbreak and provides potential ways to manage such events in the future.


The use of a fluorescent lipid as a non-covalent bound tracer of integral membrane proteins.
Solubilized and affinity purified major histocompatibility class I and class II antigens (MHC-I, MHC-II) were incorporated into liposomes, which contained a fluorescent lipid, by adsorption of the detergent to Biobeads SM-2. The fluorescent lipid was used as a non-covalently bound tracer to detect antibody-mediated agglutination of liposomes with MHC antigens. After incorporation of the membrane proteins into liposomes of different lipid composition, phosphatidylcholine was chosen as a lipid matrix. The use of a fluorescent lipid as a tracer of membrane proteins was demonstrated by the detection of human beta 2-microglobulin association with mouse MHC-I. Furthermore the method was used to measure the amount of liposomes containing two different membrane proteins.


Complexation of copper algaecide and algal organic matter in algae-laden water: Insights into complex metal-organic interactions.
Copper-based algicides have been widely used to suppress algae blooms; however, the release of algal organic matter (AOM) on account of cell lysis may cause significant changes in the mitigation, transformation, and bioavailability of Cu(II). In the present work, the binding characteristics of Cu(II) with AOM were explored via combinative characterization methods, such as high-performance size exclusion chromatography, differential absorption spectra analysis, and joint applications of two-dimensional correlation spectroscopy (2D-COS), as well as heterospectral 2D-COS and moving window 2D-COS analyses of UV, synchronous fluorescence, and FTIR spectra. Carboxyl groups displayed a preferential interaction to Cu(II) binding, followed by polysaccharides. The spectral changes of C]O stretching occur after the change of chromophores in complexation with Cu(II). The AOM chromophores exhibit obvious conformations at Cu(II) concentrations higher than 120 μM, while AOM fluorophores and functional groups exhibit the greatest changes at Cu(II) concentrations lower than 20 μM. All these observations have verified the presence of binding heterogeneity and indicate that AOM could interact with Cu(II) through diverse functional moieties. Therefore, our study contributes to the better understanding of the fate of Cu(II)-AOM complexes in aquatic systems.


Surgical treatment for brain metastases: Prognostic factors and survival in 309 patients with regard to patient age.
Brain metastases are the most common intracranial tumors. Overall, the only accepted prognostic factors are patient age and performance status. However, several other factors are considered before surgery. We performed a retrospective analysis of 309 patients who underwent surgical resection of newly diagnosed brain metastases between 1994 and 2004. Univariate survival analysis revealed age, performance status, extracranial metastases, complete resection, radiotherapy and re-craniotomy as prognostic indicators. Multivariate analysis determined that patient age, performance status, extracranial metastases, radiotherapy and re-craniotomy are independent factors of prolonged survival. We statistically estimated the age threshold separating patients with favorable outcomes from those with unfavorable prognoses. Using the Kaplan-Meier analysis this threshold can be set at 65 years. Multivariate analysis of patients >65 years revealed the presence of co-morbidities, the number of brain metastases, post-operative performance status and radiotherapy as independent prognostic factors.


Ellagic acid attenuates high-carbohydrate, high-fat diet-induced metabolic syndrome in rats.
Fruits and nuts may prevent or reverse common human health conditions such as obesity, diabetes and hypertension; together, these conditions are referred to as metabolic syndrome, an increasing problem. This study has investigated the responses to ellagic acid, present in many fruits and nuts, in a diet-induced rat model of metabolic syndrome. Eight- to nine-week-old male Wistar rats were divided into four groups for 16-week feeding with cornstarch diet (C), cornstarch diet supplemented with ellagic acid (CE), high-carbohydrate, high-fat diet (H) and high-carbohydrate, high-fat diet supplemented with ellagic acid (HE). CE and HE rats were given 0.8 g/kg ellagic acid in food from week 8 to 16 only. At the end of 16 weeks, cardiovascular, hepatic and metabolic parameters along with protein levels of Nrf2, NF-κB and CPT1 in the heart and the liver were characterised. High-carbohydrate, high-fat diet-fed rats developed cardiovascular remodelling, impaired ventricular function, impaired glucose tolerance, non-alcoholic fatty liver disease with increased protein levels of NF-κB and decreased protein levels of Nrf2 and CPT1 in the heart and the liver. Ellagic acid attenuated these diet-induced symptoms of metabolic syndrome with normalisation of protein levels of Nrf2, NF-κB and CPT1. Ellagic acid derived from nuts and fruits such as raspberries and pomegranates may provide a useful dietary supplement to decrease the characteristic changes in metabolism and in cardiac and hepatic structure and function induced by a high-carbohydrate, high-fat diet by suppressing oxidative stress and inflammation.


Physical activity and exercise improve retinal microvascular health as a biomarker of cardiovascular risk: A systematic review.
Physical activity (PA) and fitness are important modulators of vascular ageing and may therefore help expand individual health span. We aimed to systematically review the association of PA and fitness, as well as the effects of exercise interventions on the new microvascular biomarkers retinal arteriolar (CRAE) and venular (CRVE) diameters and the retinal flicker light-induced dilatation (FID) in children and adults. PubMed, Ovid, The Cochrane, EMBASE and Web of Science were searched. 805 studies were found, and 25 full-text articles analysed. Twenty-one articles were included in this systematic review. Higher PA levels were associated with narrower CRVE in children and adults. Physical inactivity was associated with wider CRVE in both age groups. Combined aerobic and motor skill training in school settings lead to wider CRAE in children. Aerobic exercise interventions in adults with or without CV risk factors induced wider CRAE and narrower CRVE. Studies on the effect of exercise on FID are scarce. In a twelve-week randomized controlled trial, high-intensity interval training significantly improved FID in older patients with CV risk factors. Higher PA and fitness levels were associated with improved retinal microvascular health in children and adults. Short-term exercise interventions in healthy children and adults, as well as CV risk patients, improved retinal microvascular structure and function. Exercise has the potential to counteract microvascular remodelling and development of small vessel disease during lifespan. Retinal vessel analysis can differentiate the beneficial effects of exercise on target microvascular organ damage.


Scleral buckling surgery for stage 4A and 4B retinopathy of prematurity in critically ill neonates.
To determine the efficacy of scleral buckling in eyes with stage 4A and 4B retinopathy of prematurity (ROP). Seven eyes of five premature infants underwent scleral buckling for stage 4 ROP in zone II. Five eyes had stage 4A ROP, and two eyes had stage 4B ROP. Six eyes had previous diode laser photocoagulation, and one eye had received an intravitreal ranibizumab injection. Scleral buckling was the procedure of choice due to lack of access to specialized pediatric vitrectomy instrumentation. Average age at surgery was 3.4 months. Postoperative anatomic retinal status, visual acuity outcome and refractive error were assessed. The scleral buckle was removed on average 8 months after surgery. Retinal reattachment was achieved in all seven eyes. At final follow-up one eye had macular ectopia and disc dragging, one eye had a macular traction fold and two eyes had optic disc pallor. Average myopic error after buckle removal was -7.5 D. Scleral buckling can be performed safely and effectively in 4A and 4B stage ROP in critically ill infants, when access to specialized pediatric vitrectomy instrumentation is limited. This surgical technique may provide adequate relief of vitreoretinal traction with improved visual potential.


Regulations and practices of genetic counselling in 38 European countries: the perspective of national representatives.
The aim of this article is to review the national regulations and practices of genetic counselling in 38 European countries, and to examine how well they intersect the ideals of genetic counselling defined in international guidelines. Using an electronic survey, representatives of the National Societies of Human Genetics in 29 countries, and appropriate contact persons for the field of genetic counselling in 9 other countries, were asked about the regulations and practices. The answers showed that consent, confidentiality, genetic counselling in the context of prenatal diagnosis, those professionals who may perform genetic counselling, and non-directiveness were the topics most often either agreed upon among professionals or regulated in those countries. These are also among the key aspects of ideal genetic counselling, based on international guidelines. Counselling in the context of susceptibility testing for multifactorial diseases, counselling people from ethnic minorities and recontacting the counsellees, on the contrary, were topics regulated or guided by generally applied practices in only few countries. Many of the answers expressed a desire for more regulation of genetic counselling, and that more uniform practices of education and organization of genetic counselling would be welcome in Europe.


Dynamic and stable hippocampal representations of social identity and reward expectation support associative social memory in male mice.
Recognizing an individual and retrieving and updating the value information assigned to the individual are fundamental abilities for establishing social relationships. To understand the neural mechanisms underlying the association between social identity and reward value, we developed Go-NoGo social discrimination paradigms that required male subject mice to distinguish between familiar mice based on their individually unique characteristics and associate them with reward availability. We found that mice could discriminate individual conspecifics through a brief nose-to-nose investigation, and this ability depended on the dorsal hippocampus. Two-photon calcium imaging revealed that dorsal CA1 hippocampal neurons represented reward expectation during social, but not non-social tasks, and these activities were maintained over days regardless of the identity of the associated mouse. Furthermore, a dynamically changing subset of hippocampal CA1 neurons discriminated between individual mice with high accuracy. Our findings suggest that the neuronal activities in CA1 provide possible neural substrates for associative social memory.


The woman who could not hold up her head.
Low serum potassium concentration is a frequently encountered abnormality, seldom accompanied by life-threatening symptoms. We present a 55-year-old women with a severe, symptomatic hypokalaemia. The pathogenesis and clinical manifestations are discussed.


Clarithromycin-loaded liposomes offering high drug loading and less irritation.
The aim of this study was to develop an efficient method of preparing less irritant clarithromycin-loaded liposomes (CLA-Lip) for injection with a high drug loading and to evaluate their physicochemical characteristics before and after lyophilization. CLA-Lip were prepared using the film-dispersion method with sodium cholesterol sulfate (SCS) and n-hexyl acid as the regulators and then lyophilized. The liposomes were characterized in terms of their size, size distribution, zeta potential, morphology, in vitro release, haemolysis, and lyophilization and irritation testing was carried out. The TEM images revealed that the structure of the CLA-Lip were multilamellar and of a regular size of around 100 nm. In addition, the lyophilized CLA-Lip were characterized by DSC and Infrared spectroscopy to confirm the structure. H-bonding and salt-forming reactions were used to ensure that clarithromycin (CLA) was stably encapsulated in the liposomes. This method provided a 30-fold increase in the concentration of clarithromycin relative to that in aqueous solution. Sucrose was found to be the best protective agent and was added in an amount of 12.5% (w/v). According to the mouse scratch test and the rat paw lick test, the pain of CLA-Lip was significantly reduce by approximately 80% compared with the solution of clarithromycin phosphate. In addition, rabbit ear vein experiments produced similar results. These findings suggested that CLA-Lip was a stable delivery system with less irritation, which should be extremely suitable for clinical application.


Challenges in pulmonary hypertension: managing the unexpected.
The diverse challenges associated with diagnosis and management of patients with pulmonary hypertension are illustrated in this case-based review. Case 1 describes a patient diagnosed with pulmonary arterial hypertension (PAH) with right heart failure and active systemic lupus erythematosus who was effectively treated with an up-front triple combination of PAH therapies and immunosuppressive therapy. In case 2, a diagnosis of pulmonary veno-occlusive disease was reached after a combined approach of clinical suspicion, physical examination, and invasive and noninvasive tests. Cautious PAH therapy and high-dose diuretics provided clinical benefit in this patient and served as a bridge to lung transplantation. These cases highlight the need for ongoing follow-up of patients with PAH, comprising frequent assessment of treatment success and continued diagnostic evaluation.


Cost analysis of three different surgical procedures for treatment of a pelvic tumour.
Malignant pelvic tumours are rare, but adequate treatment is difficult because of anatomical and functional reasons. Different surgical procedures are recommended. Besides aspects influencing the quality of life due to the different surgical techniques, costs of these surgical procedures and the perioperative time interval, depending on the chosen surgical procedure, must also be considered. Costs of three kinds of surgical treatment - internal hemipelvectomy vs external hemipelvectomy and application of an orthotic device vs ilio-femoral pseudoarthrosis - were compared, including costs of the immediate period of rehabilitation. Costs of the preoperative diagnostic procedure were excluded because they were assumed to be equal. For all calculations, treatment of a periacetabular pelvic tumour type II-b was assumed, according to the Enneking classification, with the need for a pelvic resection with wide margins including removal of the hipjoint. This analysis was performed based on the average costs per hour of physicians, nurses and physiotherapists, including a basic daily rate for additional costs, such as housekeeping and administrative costs of the hospital. In all cases, the costs of the operative procedure and the perioperative period up to 3 months were calculated, including the costs for the endoprosthetic device after internal hemipelvectomy and those for the orthotic devices after necessary external hemipelvectomy. For a postoperative period of 3 months, the cost for treatment with resection of the tumour and performing an ilio-femoral pseudoarthrosis is nearly DM 56,741.54; treatment with an internal hemipelvectomy including the endoprosthetic pelvic replacement costs DM 81,439.34; and treatment with external hemipelvectomy and application of an orthotic device with a pelvic cage costs DM 69,138.46. In this analysis, social costs due to the different rates of disability for years or costs of a new prosthetic device after years and aspects related to resulting quality of life were excluded. In contrast, costs of three different actual cases can differ significantly from the estimated costs. The most important influencing items are surgical implants, blood units and similar products, and surgical treatment of postoperative complications. Regarding all these aspects, limb salvage seems to be advantageous over amputation. Whether an ileo-femoral pseudoarthrosis or endoprosthetic pelvic replacement should be performed is a decision that should be made by the physicians. based on the underlying diagnosis and the correlated expected survival. Furthermore, additional factors probably reducing the rate of postoperative complications, such as infections or necrosis following radiotherapy and being summarised under the term "quality of life", should also be regarded.


Radioaffinity assay, a new approach to binding assays applied to the measurement of serum albumin.
The affinity adsorbent Cibacron Blue F3GA-Sepharose 4-B has been used to develop a binding assay for human serum albumin. The method is simple, accurate and precise. It agrees well with the 'rocket' immunoelectrophoretic method and is proposed as an alternative technique for the re-estimation of albumin levels in the low range (less than 30 g/l) where the bromocresol green dye-binding method overestimates. Bilirubin, several drugs, gammaglobulin, haemoglobin and heparin do not interfere with the estimations.


Cerebrospinal fluid myelin basic protein is elevated in multiple system atrophy.
Parkinson's disease (PD) and multiple system atrophy (MSA) have overlapping symptoms, challenging an early diagnosis. Diagnostic accuracy is important because PD and MSA have a different prognosis and response to treatment. Here, we aimed to evaluate the diagnostic value of brain-specific structural proteins in cerebrospinal fluid (CSF) of PD and MSA patients, as well as their association with cognitive decline. CSF samples were collected from patients with clear signs of parkinsonism, but with uncertain diagnosis at the time of inclusion. Clinical diagnoses of PD (n = 55) and MSA (n = 22) were established after 3 and 10 years of follow-up and re-evaluated after 12 years, according to the most updated clinical criteria. CSF from controls (n = 118) was studied for comparison. Neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP), S100 calcium-binding protein B (S100B) and myelin basic protein (MBP) levels in CSF were measured using ELISA. Protein levels were also correlated with cognitive decline, i.e. worsening of the mini mental state examination (MMSE) over a period of three years. MBP concentrations were increased in MSA compared to PD and controls (p < 0.005) and could differentiate MSA and PD with high accuracy (AUC = 0.781; p < 0.001). Concentrations of MPB, GFAP and S100B, but not NSE, were significantly elevated in PD patients compared to controls (p = 0.05). None of the brain-specific structural proteins correlated with MMSE progression. Our results demonstrate that MBP differentiates PD from MSA at early stages of the disease, indicating that demyelination and axonal damage may already occur in early stages of MSA.


Circuit-guided population acclimation of a synthetic microbial consortium for improved biochemical production.
Microbial consortia have been considered potential platforms for bioprocessing applications. However, the complexity in process control owing to the use of multiple strains necessitates the use of an efficient population control strategy. Herein, we report circuit-guided synthetic acclimation as a strategy to improve biochemical production by a microbial consortium. We designed a consortium comprising alginate-utilizing Vibrio sp. dhg and 3-hydroxypropionic acid (3-HP)-producing Escherichia coli strains for the direct conversion of alginate to 3-HP. We introduced a genetic circuit, named "Population guider", in the E. coli strain, which degrades ampicillin only when 3-HP is produced. In the presence of ampicillin as a selection pressure, the consortium was successfully acclimated for increased 3-HP production by 4.3-fold compared to that by a simple co-culturing consortium during a 48-h fermentation. We believe this concept is a useful strategy for the development of robust consortium-based bioprocesses.


Origin and evolutionary malleability of T cell receptor α diversity.
Lymphocytes of vertebrate adaptive immune systems acquired the capability to assemble, from split genes in the germline, billions of functional antigen receptors1-3. These receptors show specificity; unlike the broadly tuned receptors of the innate system, antibodies (Ig) expressed by B cells, for instance, can accurately distinguish between the two enantiomers of organic acids4, whereas T cell receptors (TCRs) reliably recognize single amino acid replacements in their peptide antigens5. In developing lymphocytes, antigen receptor genes are assembled from a comparatively small set of germline-encoded genetic elements in a process referred to as V(D)J recombination6,7. Potential self-reactivity of some antigen receptors arising from the quasi-random somatic diversification is suppressed by several robust control mechanisms8-12. For decades, scientists have puzzled over the evolutionary origin of somatically diversifying antigen receptors13-16. It has remained unclear how, at the inception of this mechanism, immunologically beneficial expanded receptor diversity was traded against the emerging risk of destructive self-recognition. Here we explore the hypothesis that in early vertebrates, sequence microhomologies marking the ends of recombining elements became the crucial targets of selection determining the outcome of non-homologous end joining-based repair of DNA double-strand breaks generated during RAG-mediated recombination. We find that, across the main clades of jawed vertebrates, TCRα repertoire diversity is best explained by species-specific extents of such sequence microhomologies. Thus, selection of germline sequence composition of rearranging elements emerges as a major factor determining the degree of diversity of somatically generated antigen receptors.


Oxidative stress and phthalate-induced down-regulation of steroidogenesis in MA-10 Leydig cells.
Previous studies have shown that phthalate exposure can suppress steroidogenesis. However, the affected components of the steroidogenic pathway, and the mechanisms involved, remain uncertain. We show that incubating MA-10 Leydig cells with mono-(2-ethylhexyl) phthalate (MEHP) resulted in reductions in luteinizing hormone (LH)-stimulated cAMP and progesterone productions. cAMP did not decrease in response to MEHP when the cells were incubated with cholera toxin or forskolin. Incubation of MEHP-treated cells with dibutyryl-cAMP, 22-hydroxycholesterol or pregnenolone inhibited the reductions in progesterone. Increased levels of reactive oxygen species (ROS) occurred in response to MEHP. In cells in which intracellular glutathione was depleted by buthionine sulfoximine pretreatment, the increases in ROS and decreases in progesterone in response to MEHP treatment were exacerbated. These results indicate that MEHP inhibits MA-10 Leydig cell steroidogenesis by targeting LH-stimulated cAMP production and cholesterol transport, and that a likely mechanism by which MEHP acts is through increased oxidative stress.


Detecting protein-small molecule interactions using fluorous small-molecule microarrays.
General binding assays involving microarrays of small molecules can be used to identify small molecule ligands for nearly any protein, even in the absence of knowledge about protein structure or function. Several suitable methods for manufacturing small molecule microarrays (SMMs) exist and different immobilization methods may be more or less preferable for any given application. Here, we describe a protocol for noncovalent and homogenous capture of small molecules using fluorous interactions between small molecules containing fluorocarbon tags and fluorocarbon-coated glass surfaces. These arrays are especially useful for applications that require display of compounds in a specific orientation such as screening biased libraries.


Mitosis: new roles for myosin-X and actin at the spindle.
Roles for actin and myosin in positioning mitotic spindles in the cell are well established. A recent study of myosin-X function in early Xenopus embryo mitosis now reports that this unconventional myosin is required for pole integrity and normal spindle length by localizing to poles and exerting pulling forces on actin filaments within the spindle.


Correction to: Optimality criteria for futility stopping boundaries for group sequential designs with a continuous endpoint.
An amendment to this paper has been published and can be accessed via the original article.


Caruncle dysgeneses - A case series.
Caruncle dysgeneses are extremely rare and must be differentiated from caruncular and conjunctival tumors. Very few case reports with histopathological descriptions exist. In this case series, four patients with five caruncle dysgeneses, two with histopathological findings, are characterised. Patient 1, a 26-year-old woman, presented with a conjunctival change at the left lower eyelid she had first noticed seven months earlier. She reported foreign body sensation and itching. On her left eye was a subtarsal conjunctival tumour measuring approximately 4 × 4 mm with whitish sebaceous gland-like inclusions located almost in the fornix morphologically resembling the nearby caruncle. The patient was asymptomatic after excision. Histopathological examination of the excised tissue showed non-keratinizing squamous epithelium with goblet cells. Subepithelially, there was lymphoplasmacytic cellular infiltration with intervening epidermal cysts adjacent to sebaceous glands and underlying adipose tissue, but no hair follicles or sweat/lacrimal glands. The epidermal cysts contained scattered hairs. A diagnosis of supernumerary caruncle was made.Patient 2, a 56-year-old woman, was referred for evaluation of a caruncle tumour that was reported to be present since childhood. Clinically the 5 × 5 mm measuring tumour appeared yellowish and less reflective compared to the normal caruncle tissue. Histopathologically, non-keratinizing squamous epithelium with goblet cells was found. In the area of more exposed tumour tissue, there were significantly fewer goblet cells and incipient keratinization of the superficial epithelial layers. Subepithelially, sebaceous glands and adipocytes were present. Hair follicles or sweat/lacrimal glands were not evident. A diagnosis of megacaruncle was made.Patient 3, a 58-year-old woman with Goldenhar syndrome, was clinically diagnosed with a supernumerary caruncle on the right eye as an incidental finding.Patient 4, a 24-year old man, clinically presented with a megacaruncle on the right eye and a supernumerary caruncle on the left. Caruncle dysgeneses are often asymptomatic and have to be differentiated from other caruncular and conjunctival tumors. If they are present, attention should be paid to signs of an oculo-auriculo-vertebral spectrum as Goldenhar syndrome. In case of unclear findings or complaints, excision with subsequent histopathological examination is required.


Unsupervised representation learning improves genomic discovery and risk prediction for respiratory and circulatory functions and diseases.
High-dimensional clinical data are becoming more accessible in biobank-scale datasets. However, effectively utilizing high-dimensional clinical data for genetic discovery remains challenging. Here we introduce a general deep learning-based framework, REpresentation learning for Genetic discovery on Low-dimensional Embeddings (REGLE), for discovering associations between genetic variants and high-dimensional clinical data. REGLE uses convolutional variational autoencoders to compute a non-linear, low-dimensional, disentangled embedding of the data with highly heritable individual components. REGLE can incorporate expert-defined or clinical features and provides a framework to create accurate disease-specific polygenic risk scores (PRS) in datasets which have minimal expert phenotyping. We apply REGLE to both respiratory and circulatory systems: spirograms which measure lung function and photoplethysmograms (PPG) which measure blood volume changes. Genome-wide association studies on REGLE embeddings identify more genome-wide significant loci than existing methods and replicate known loci for both spirograms and PPG, demonstrating the generality of the framework. Furthermore, these embeddings are associated with overall survival. Finally, we construct a set of PRSs that improve predictive performance of asthma, chronic obstructive pulmonary disease, hypertension, and systolic blood pressure in multiple biobanks. Thus, REGLE embeddings can quantify clinically relevant features that are not currently captured in a standardized or automated way.


Impact of providing milk options and nutrient information in school lunch progams.
The impact of offering junior and senior high school students a choice of whole, skim, and low-fat milk within the Type A lunch pattern, with and without providing nutritional information at point of purchase, was investigated. When such an option was offered, approximately 13 per cent of the students chose either skim or low-fat milk. When nutritional information was made available concurrently, this percentage increased to 20, with more than 80 per cent of those students choosing low-fat milk. The older students used the options significantly more frequently than did the junior high students. Waste of skim milk, the taste of which was described as watery, was greater than that of the other types of milk. Although most students chose the type of milk they had at home, 16 per cent indicated that their choice was influenced by information on the fat and caloric content of milks provided by posters.


Pressure and tendon strain in the sixth extensor compartment of the wrist during simulated provocative maneuvers for diagnosing extensor carpi ulnaris tendinitis.
Various provocative maneuvers for diagnosing extensor carpi ulnaris (ECU) tendinitis have been reported; however, it remains unclear which maneuver is the most sensitive to detect ECU tendinitis. To clarify this, we investigated and compared the extratendinous pressure and ECU tendon strain in the sixth extensor compartment of the wrist during various provocative maneuvers for diagnosing ECU tendinitis. Nine upper extremities from nine fresh-frozen cadavers were examined. We investigated extratendinous pressure in the ECU fibro-osseous tunnel of the distal ulna and ECU tendon strain during eight forearm positions-neutral rotation, pronation, supination, pronation with wrist flexion, supination with wrist flexion, supination with wrist extension, both hand and forearm supination, and supination with ECU full loading-to simulate provocative maneuvers reported to detect ECU tendinitis. Pressure was significantly higher during both hand and forearm supination (carpal supination test) and during supination with wrist extension (prayer's hand supination test) than during neutral rotation. The pressure during the carpal supination test was 3 times higher than that during the prayer's hand supination test and 27 times higher than that during the neutral position. Strain was significantly higher during the carpal supination test and during supination with ECU full loading (the ECU synergy test) than during other maneuvers. Both pressure and tendon strain increased most notably during the carpal supination test compared to the other maneuvers, which suggests that the carpal supination test is the most sensitive for the detection of ECU tendinitis.


Comparison of clinical and radiological outcomes after three different surgical treatments for resistant calcifying tendinitis of the shoulder: a short-term randomized controlled trial.
A preferable surgical treatment for patients with conservative therapy-resistant calcifying tendinitis of the shoulder is still a matter of debate. Therefore, the purpose of this study was to evaluate and compare short-term clinical and radiological results of three surgical treatment options for these patients. A multicenter randomized trial was conducted. Sixty-nine patients were randomly assigned to receive 1. subacromial decompression (Group SAD), 2. debridement of calcifications (Group D), or 3. debridement of calcifications with SAD (Group D + SAD). Stringent inclusion and exclusion criteria were used. The primary outcome was an improvement in VAS for pain (pVAS) 6 months postoperatively. Secondary outcomes were an improvement in pVAS 6 weeks postoperatively, functional outcomes (CMS, DASH, ASES), radiological outcome, additional treatments, and complications. The improvement in pVAS was significant in all groups (p < 0.001) and did not differ between the groups after 6 months. Six weeks postoperatively, the improvement in pVAS was significantly (p = 0.03) less in Group SAD compared to Group D + SAD (16.5 mm, SD 19.3 mm vs 33.1 mm, SD 19.7 mm, respectively). The mean size of calcifications decreased significantly in all groups (p < 0.0001). In Group SAD, the size of the calcifications decreased less (p = 0.04) compared to Group D and Group D + SAD after 6 weeks. Group SAD received more additional treatments (p = 0.003) compared to Group D + SAD (9 vs 1), which were mainly subacromial cortisone injections. All patient groups showed significant pain relief and an improvement in shoulder function 6 months after surgery. However, patients in Group SAD showed inferior pain relief and less improvement in DASH score after 6 weeks. Furthermore, this group required more postoperative additional treatments. No significant differences in clinical and radiological outcomes were observed between patients in Group D compared to Group D + SAD. Therefore, an arthroscopic debridement without subacromial decompression seems to be advisable for patients with therapy-resistant calcifying tendinitis of the shoulder. Level of evidence 2, Open-Label Randomized Clinical Trial. IRB METC Zuyderland MC. Number: 14-T-112. Registered at trialregister.nl NL 4947.


Emerging mass spectrometry-based approaches to probe protein-receptor interactions: focus on overcoming physiological barriers.
Physiological barriers, such as the blood-brain barrier and intestinal epithelial barrier, remain significant obstacles towards wider utilization of biopharmaceutical products. Receptor-mediated transcytosis has long been viewed as an attractive means of crossing such barriers, but successful exploitation of this route requires better understanding of the interactions between the receptors and protein-based therapeutics. Detailed characterization of such processes at the molecular level is challenging due to the very large physical size and heterogeneity of these species, which makes use of many state-of-the art analytical techniques, such as high-resolution NMR and X-ray crystallography impractical. Mass spectrometry has emerged in the past decade as a powerful tool to study protein-receptor interactions, although its applications to investigate interaction of biopharmaceuticals with their physiological partners are still limited. We highlight the potential of this technique by considering several recent examples where it had been instrumental for understanding molecular mechanisms critical for receptor-mediated transcytosis of transferrin-based therapeutics.


Systematic review of attitudes toward donation after cardiac death among healthcare providers and the general public.
Organ donation after cardiac death (DCD) is one promising possibility of combating the organ shortage, but it raises ethical issues that differ from those raised in donation after brain death (DBD). Also, DCD may be perceived differently than DBD by medical staff and the public. The aim of this article is to systematically review empirical studies on attitudes of medical personnel and the public toward DCD and to discuss the findings from an ethical perspective. Our study was conducted in accordance with a seven-step approach for systematic reviews of empirical studies in bioethics. The authors chose PubMed, EMBASE, CINAHL, PSYCINFO, and PSYNDEX, thus attempting to cover biomedical, sociological and ethical articles on the subject. A search algorithm using controlled vocabulary of the respective databases (where applicable) was created, and criteria for the relevance assessment of the articles were established. Article quality was assessed using the Critical Appraisal Skills Programme tool. The authors took an integrative approach to the data, combining it for further analysis. Qualitative data were synthesized by means of thematic analysis, and a spectrum of relevant themes was identified. Then the authors extracted the quantitative data that corresponded with the identified themes. Quantitative data on common subjects were juxtaposed and presented later. Identified themes were the levels of support for DBD vs. DCD, attitudes toward postmortem measures without previous consent, lack of knowledge about DCD, concerns about the Dead Donor Rule, the potential for conflict of interest, making donation happen, and the call for standardized DCD protocols. All of these issues are of ethical relevance and merit further discussion. We conclude that deep-rooted concerns about DCD exist among medical personnel and the general public. These need to be taken seriously in order to maintain or foster trust in the transplantation system.


Sagebrush and grasshopper responses to atmospheric carbon dioxide concentration.
Seed- and clonally-propagated plants of Big Sagebrush (Artemisia tridentata var.tridentata) were grown under atmospheric carbon dioxide regimes of 270, 350 and 650 μl l-1 and fed toMelanoplus differentialis andM. sanguinipes grasshoppers. Total shrub biomass significantly increased as carbon dioxide levels increased, as did the weight and area of individual leaves. Plants grown from seed collected in a single population exhibited a 3-5 fold variation in the concentration of leaf volatile mono- and sesquiterpenes, guaianolide sesquiterpene lactones, coumarins and flavones within each CO2 treatment. The concentration of leaf allelochemicals did not differ significantly among CO2 treatments for these seed-propagated plants. Further, when genotypic variation was controlled by vegetative propagation, allelochemical concentrations also did not differ among carbon dioxide treatments. On the other hand, overall leaf nitrogen concentration declined significantly with elevated CO2. Carbon accumulation was seen to dilute leaf nitrogen as the balance of leaf carbon versus nitrogen progressively increased as CO2 growth concentration increased. Grasshopper feeding was highest on sagebrush leaves grown under 270 and 650 μl l-1 CO2, but varied widely within treatments. Leaf nitrogen concentration was an important positive factor in grasshopper relative growth but had no overall effect on consumption. Potential compensatory consumption by these generalist grasshoppers was apparently limited by the sagebrush allelochemicals. Insects with a greater ability to feed on chemically defended host plants under carbon dioxide enrichment may ultimately consume leaves with a lower nitrogen concentration but the same concentration of allelochemicals. Compensatory feeding may potentially increase the amount of dietary allelochemicals ingested for each unit of nitrogen consumed.


Vedolizumab as induction and maintenance therapy for Crohn's disease.
The efficacy of vedolizumab, an α4β7 integrin antibody, in Crohn's disease is unknown. In an integrated study with separate induction and maintenance trials, we assessed intravenous vedolizumab therapy (300 mg) in adults with active Crohn's disease. In the induction trial, 368 patients were randomly assigned to receive vedolizumab or placebo at weeks 0 and 2 (cohort 1), and 747 patients received open-label vedolizumab at weeks 0 and 2 (cohort 2); disease status was assessed at week 6. In the maintenance trial, 461 patients who had had a response to vedolizumab were randomly assigned to receive placebo or vedolizumab every 8 or 4 weeks until week 52. At week 6, a total of 14.5% of the patients in cohort 1 who received vedolizumab and 6.8% who received placebo were in clinical remission (i.e., had a score on the Crohn's Disease Activity Index [CDAI] of ≤150, with scores ranging from 0 to approximately 600 and higher scores indicating greater disease activity) (P=0.02); a total of 31.4% and 25.7% of the patients, respectively, had a CDAI-100 response (≥100-point decrease in the CDAI score) (P=0.23). Among patients in cohorts 1 and 2 who had a response to induction therapy, 39.0% and 36.4% of those assigned to vedolizumab every 8 weeks and every 4 weeks, respectively, were in clinical remission at week 52, as compared with 21.6% assigned to placebo (P<0.001 and P=0.004 for the two vedolizumab groups, respectively, vs. placebo). Antibodies against vedolizumab developed in 4.0% of the patients. Nasopharyngitis occurred more frequently, and headache and abdominal pain less frequently, in patients receiving vedolizumab than in patients receiving placebo. Vedolizumab, as compared with placebo, was associated with a higher rate of serious adverse events (24.4% vs. 15.3%), infections (44.1% vs. 40.2%), and serious infections (5.5% vs. 3.0%). Vedolizumab-treated patients with active Crohn's disease were more likely than patients receiving placebo to have a remission, but not a CDAI-100 response, at week 6; patients with a response to induction therapy who continued to receive vedolizumab (rather than switching to placebo) were more likely to be in remission at week 52. Adverse events were more common with vedolizumab. (Funded by Millennium Pharmaceuticals; GEMINI 2 ClinicalTrials.gov number, NCT00783692.).


Association between self-reported race and ethnicity and myositis-specific autoantibodies in a diverse cohort of patients with inflammatory myopathy.
Myositis-specific autoantibodies (MSAs) are highly specific biomarkers for idiopathic inflammatory myopathies (IIMs). We investigated whether self-reported race and ethnicity were associated with the presence of specific MSAs. Charts of patients with IIM seen at 3 large healthcare systems in the same US city were reviewed. Demographic data and MSA test results were abstracted. Associations between race and ethnicity and presence of MSAs were analyzed using bivariate analysis and further characterized using separate unadjusted and adjusted logistic regression models. One hundred twenty-one subjects were included (19% Asian, 10% Black or African American, 27% Latinx or Hispanic, 36% non-Hispanic White, and 7% Other). In a bivariate analysis, anti-Jo-1 and anti-MDA5 autoantibodies were associated with race and ethnicity (p = 0.03 and 0.02, respectively). Black or African American subjects had increased odds of a positive anti-Jo-1 result compared to non-Hispanic White subjects on unadjusted logistic regression analysis (OR 8.61, 95% CI 1.61-46.07), although after adjustment for age and gender this finding was not significant. Subjects categorized as Other had increased odds of a positive anti-MDA5 result compared to non-Hispanic White subjects on both unadjusted (OR 55.0, 95% CI 2.02-1493) and adjusted analyses (OR 44.8, 95% CI 1.55-1298). Anti-Jo-1 and anti-MDA5 autoantibodies were significantly associated with race and ethnicity on bivariate analysis. Black or African American subjects had increased odds of positive anti-Jo-1 autoantibody on unadjusted, but not adjusted, logistic regression analysis. Subjects characterized as Other had increased odds of positive anti-MDA5 autoantibody, although confidence intervals were wide. Key Points • Association found between MSAs and race and ethnicity in diverse US cohort • Anti-Jo-1 and anti-MDA5 associated with race and ethnicity in bivariate analyses.


Do socio-demographic characteristics and/or health status explain the magnitude of differences between patient and general public utility values? A chronic low back pain patients case study.
Utility values can be obtained from different respondent groups, including patients and members of the general public. Evidence suggests that patient values are typically higher than general public values. This study explores whether the magnitude of disagreement between both values can be explained by socio-demographic characteristics and/or health status. Data of 5037 chronic low back pain patients were used. Self-reported EQ-VAS was employed as a proxy of patients' preference for their own health state. General public values for the patients' EQ-5D-3L health states were obtained using the Dutch VAS-based tariff. The difference between patient and general public values was assessed using a paired t-test. Subsequently, this difference was used as a dependent variable and regressed upon dummy variables of socio-demographic and health status characteristics. Coefficients represented age, gender, education level, social support, back pain intensity, leg pain intensity, functional status, comorbidities, catastrophizing, and treatment expectations. Patient values were higher than general public values (0.069; 95%CI:0.063-0.076). The magnitude of disagreement between both values was associated with age, gender, education level, social support, functional status, and comorbidities, but not with back pain intensity, leg pain intensity, catastrophizing, and treatment expectations. Patients were found to value their own health status higher than members of the general public. The magnitude of disagreement between both values was found to differ by various socio-demographic and/or health status characteristics. This suggest that patient characteristics account for a relevant fraction of the identified disagreements between patient and general public values, and that mechanisms thought to be responsible for these disagreements, such as adaptation and response shift, have a differential impact across patient sub-groups.


Transfer of experimental autoimmune thyroiditis to normal syngeneic mice by injection of mouse thyroglobulin-sensitized T lymphocytes after activation with concanavalin A.
Induction of autoimmune thyroiditis in normal syngeneic CBA/J mice was achieved by injection of 72-hr concanavalin A (Con A)-induced lymphoblasts from donor mice which had been immunized with mouse thyroglobulin (MTg) emulsified with complete Freund's adjuvant (CFA). Injection of lymph node or spleen cells, or frequent injection of serum taken from mice with autoimmune thyroiditis failed to transfer appreciable thyroiditis to recipient mice. Selection by treatment of incubated cells with monoclonal antibody and complement revealed that effector cells in Con A-induced lymphoblast populations for the transfer of autoimmune thyroiditis were Thy-1.2+, Lyt-1.1+, and Lyt-2.1- lymphocytes. These results demonstrate that experimental autoimmune thyroiditis can be adoptively transferred into naive mice by activated Thy-1+, Lyt-2- lymphoblasts.


Relationship between hepatic glutathione content and carbon tetrachloride-induced hepatotoxicity in vivo.
The relationship between carbon tetrachloride (CCl4)-induced hepatotoxicity and hepatic glutathione (GSH) content was investigated in fed and fasted rats. The elevation of serum glutamic-pyruvic transaminase (GTP) activity by CCl4 treatment was enhanced by fasting. Although the hepatic GSH content fo 12-hour-fasted rats was higher than that of fed rats determined at 6 p.m., the serum GPT activity of the former was higher than that of the latter. Starvation had no effect on the activities of hepatic glutathione peroxidase (GSH-Px) and glutathione reductase (GR). The results suggest that the potentiation of hepatic injury by CCl4 cannot be related to hepatic GSH content.


Effective and rapid technique for temporal response modeling of surface acoustic wave interdigital transducers.
Surface Acoustic Wave Interdigital Transducers (SAW-IDT) has a considerable application potential for characterization of properties of thin layers, coatings and functional surfaces. For optimization of these SAW-IDTs, it is necessary to study various SAW-IDT configurations by varying the number of electrodes, dimensions of the electrodes, their shapes and spacings. The finite element method (FEM) is generally used to model such transducers but results are obtained in several hours (or days). Thus it is necessary to implement effective and rapid technique for SAW-IDT modeling. In this study, we develop simulation tool based on Spatial Impulse Response model. Therefore, we reduce considerably computing time and results are obtained in a few seconds. In order to validate this method, theoretical and experimental results are compared with finite element method. The results obtained show a good concordance and confirm effectiveness of suggested method. In additional, this method requires less computer memory.


Neomycin is a platelet-derived growth factor (PDGF) antagonist that allows discrimination of PDGF alpha- and beta-receptor signals in cells expressing both receptor types.
The aminoglycoside neomycin has recently been found to affect certain platelet-derived growth factor (PDGF) responses in C3H/10T1/2 C18 fibroblasts. Using porcine aortic endothelial cells transfected with PDGF alpha- or beta-receptors, we explored the possibility that neomycin interferes with the interaction between the different PDGF isoforms and their receptors. We found that neomycin (5 mM) inhibited the binding of 125I-PDGF-BB to the alpha-receptor with only partial effect on the binding of 125I-PDGF-AA; in contrast, the binding of 125I-PDGF-BB to the beta-receptor was not affected by the aminoglycoside. Scatchard analyses showed that neomycin (5 mM) decreased the number of binding sites for PDGF-BB on alpha-receptor-expressing cells by 87%. Together with cross-competition studies with 125I-labeled PDGF homodimers, the effect of neomycin indicates that PDGF-AA and PDGF-BB bind to both common and unique structures on the PDGF alpha-receptor. Neomycin specifically inhibited the autophosphorylation of the alpha-receptor by PDGF-BB, with less effect on the phosphorylation induced by PDGF-AA and no effect on the phosphorylation of the beta-receptor by PDGF-BB. Thus, neomycin is a PDGF isoform- and receptor-specific antagonist that provides a possibility to compare the signal transduction pathways of alpha- and beta-receptors in cells expressing both receptor types. This approach was used to show that activation of PDGF beta-receptors by PDGF-BB mediated a chemotactic response in human fibroblasts, whereas activation of alpha-receptors by the same ligand inhibited chemotaxis.


Oxidative stress induced by the chemotherapeutic agent arsenic trioxide.
Arsenic compounds have been used for medicinal purposes throughout history. Arsenic trioxide (As2O3) achieved dramatic remissions in patients with acute promyelocytic leukaemia. Unfortunately, the clinical usefulness of As2O3 has been limited by its toxicity. The present study was designed to investigate the toxic effects of As2O3 at its clinical concentrations. Experimental rats were administered with As2O3 2, 4 and 8 mg/kg body weight for a period of 45 days and the serum glucose, creatine kinase, lactate dehydrogenase, lipid peroxidation and antioxidant status were measured. As2O3-treated rats showed elevated serum glucose, creatine kinase and lactate dehydrogenase concentrations. Lipid peroxidation product malondialdehyde was found to be produced more in arsenic-treated rats. Reduced glutathione and glutathione-dependant antioxidant enzymes, glutathione-S-transferase and glutathione peroxidase, and the antiperoxidative enzymes, superoxide dismutase and catalase, concentrations were reduced with the As2O3 treatment. All these toxic effects were found increased with the increase in concentration of As2O3. The results of the study indicate that As2O3 produced dose-dependant toxic side effects at its clinical concentrations.


tele-Methylhistamine levels and histamine turnover in nuclei of the rat hypothalamus and amygdala.
An HPLC method using fluorescence detection for the determination of tele-methylhistamine (t-MH) was improved to a sensitivity level which enabled the detection of 0.05 pmol of tissue t-MH. The t-MH contents and the histamine turnover rates in various nuclei of the rat hypothalamus and amygdala were subsequently measured. The histamine turnover rates were estimated from pargyline-induced t-MH accumulation. Both the t-MH levels and the histamine turnover rates were shown to be relatively high in the nuclei dorsomedialis and premammillaris ventralis of the hypothalamus, and also in the nucleus medialis of the amygdala. The steady-state t-MH levels in various nuclei of the hypothalamus and amygdala correlated well with the histamine turnover rates in these nuclei.


Characterization of input synapses on intracellularly stained neurons in hippocampal slices: an HRP/EM study.
This study describes the fine structure of input synapses on identified neurons in slices of the guinea pig hippocampus. For morphological identification, granule cells of the fascia dentata and pyramidal neurons of regio inferior of the hippocampus were impaled and intracellularly stained with horseradish peroxidase (HRP). Input synapses on the HRP-stained neurons were identified in the electron microscope by the location of the synapses in inner or outer zones of the dentate molecular layer, as in the case of the synaptic contacts on injected granule cells, or by unique fine structural characteristics, as in the case of the giant mossy fiber boutons on CA3 pyramidal cells. As in tissue fixed in situ by transcardial perfusion, a large number of terminals arising from the different afferents in inner and outer zones of the dentate molecular layer were well preserved and formed synaptic contacts with small spines, large complex spines, and dendritic shafts of the HRP-filled granule cells. Mossy fiber synapses on the stained CA3 neurons were densely filled with clear vesicles, contained a few dense-core vesicles, and formed synaptic contacts with large spines or excrescences. Occasionally electrondense degenerating boutons were also found impinging on the stained dendrites and spines. The significance of the present findings for electrophysiological and pharmacological studies on brain slices is discussed.


Taxonomy of Leucochloridium sp. (Digenea) infecting Succinea pfeifferi Rossmässler, 1835.
The anatomical and morphological variability of sporocysts and metacercariae of Leucochloridium sp. recovered from natural infected Succinea pfeifferi Rossmässler collected in the Agdenes area. Norway (63 degrees 35'N, 9 degrees 45'E), was studied by light microscopy. The results are compared with five species from the Nearctic and Palearctic with known adult stage (L. variae McIntosh, 1932; L. fuscostriatum Robinson, 1947; L. perturbatum Pojmanska, 1969; L. subtilis Pojmanska, 1969; L. fuscum Rietschel, 1970) and nine species with unknown adult stage. In the larval stages no morphological taxonomical characters were found to separate Leucochloridium sp. as a species distinct from the five (and nine) species. The variability and validity of the characters used is discussed.


CTNNB1 p.D32A (c.95A > C) somatic mutation in stage I grade 1 endometrioid endometrial carcinoma with lung metastasis: a case report.
Most endometrial cancers are of low histological grade and uterine-confined, with a high 5-year survival rate. However, a small subset of women with low-grade and early-stage endometrioid endometrial cancer experience recurrence and death; thus, a more precise risk-stratification is needed. A 29-year-old woman presented with abnormal vaginal bleeding and was diagnosed with FIGO grade 1 endometrioid endometrial carcinoma by curettage. Comprehensive cancer staging including pelvic and para-aortic lymphadenectomy was then performed. Postoperative pathological findings suggested an FIGO grade 1 endometrioid endometrial carcinoma infiltrating the superficial muscle layer. The patient did not receive adjuvant therapy. After 4 years of follow-up, the patient returned to our institution with lung metastasis. She underwent thoracoscopic resection of the affected lobes, followed by six cycles of combined chemotherapy of paclitaxel and carboplatin. Next-generation sequencing showed that the primary and lung metastatic tumors shared 4 mutations: PTEN (p.P248Lfs*8), CTNNB1 (p.D32A), BCOR (p.N1425S) and CBL (p.S439N). Immunohistochemistry revealed nuclear location of β-catenin in the primary and lung metastatic tumor samples, indicating abnormal activation of β-catenin. CTNNB1p.D32A (c.95A > C) mutation may be related to lung metastasis in this patient with low-grade early-stage endometrioid endometrial carcinoma.


Role of lymphangiogenic factors in tumor metastasis.
Nearly four centuries after the discovery of lymphatic vessels, the molecular mechanisms underlying their development are beginning to be elucidated. Vascular endothelial growth factor C (VEGF-C) and VEGF-D, via signaling through VEGFR-3, appear to be essential for lymphatic vessel growth. Observations from clinicopathological studies have suggested that lymphatic vessels serve as the primary route for the metastatic spread of tumor cells to regional lymph nodes. Recent studies in animal models have provided convincing evidence that tumor lymphangiogenesis facilitates lymphatic metastasis. However, it is not clear how tumor-associated lymphangiogenesis is regulated, and little is known about how tumor cells escape from the primary tumor and gain entry into the lymphatics. This review examines some of these issues and provides a brief summary of the recent developments in this field of research.


Truncated Thioredoxin Peptides Serves as an Efficient Fusion Tag for Production of Proinsulin.
Insulin is a peptide hormone used for regulating blood glucose levels. Human insulin market is projected to grow at a rate of 12.5% annually. To meet the needs of patients, a cost effective insulin manufacturing strategy has to be developed. This can be achieved by selecting a competent host, ideal fusion tag and streamlined downstream process. In this article, we have demonstrated that selecting a right fusion partner for expression of toxic proteins like insulin, plays a major role in increasing the recombinant protein yield. In this article, we have focused on identifying a peptide tag fusion partner for expressing proinsulin by truncating thioredoxin tag. Truncations were carried out from both Amino and Carboxy terminus of the protein and efficiency of truncated sequences was evaluated by expressing it with proinsulin gene. FCTRX (1-15) sequence fused to proinsulin was processed further to establish downstream protocol for purification. Thioredoxin tag was truncated appropriately by considering the fusion tag: protein ratio. A couple of sequences ranging 10 - 15 amino acids were identified based on its in silico properties. Of these FCTRX (1-15) showed increased expression and stability of fusion protein. 156 mg of purified insulin was generated from 1g of inclusion body after enzymatic conversion and chromatographic steps. As a result of the current study, it was concluded that FCTRX (1-15) peptide has advantageous attributes to be considered as an ideal fusion tag for expression of proinsulin. This can be further explored by expressing it with other proteins.


Characterization of corrosion scale formed on stainless steel delivery pipe for reclaimed water treatment.
To reveal corrosion behavior of stainless steel delivery pipe used in reclaimed water treatment, this research focused on the morphological, mineralogical and chemical characteristics of stainless steel corrosion scale and corroded passive film. Corrosion scale and coupon samples were taken from a type 304 pipe delivering reclaimed water to a clear well in service for more than 12 years. Stainless steel corrosion scales and four representative pipe coupons were investigated using mineralogy and material science research methods. The results showed corrosion scale was predominantly composed of goethite, lepidocrocite, hematite, magnetite, ferrous oxide, siderite, chrome green and chromite, the same as that of corroded pipe coupons. Hence, corrosion scale can be identified as podiform chromite deposit. The loss of chromium in passive film is a critical phenomenon when stainless steel passive film is damaged by localized corrosion. This may provide key insights toward improving a better comprehension of the formation of stainless steel corrosion scale and the process of localized corrosion. The localized corrosion behavior of stainless steel is directly connected with reclaimed water quality parameters such as residual chlorine, DO, Cl(-) and SO4(2-). In particular, when a certain amount of residual chlorine in reclaimed water is present as an oxidant, ferric iron is the main chemical state of iron minerals.


Urine cytology in renal glomerular disease and value of G1 cell in the diagnosis of glomerular bleeding.
The objectives of the present study were to evaluate the cytology of urine sediments in patients with glomerular diseases, as well as the value of G1 dysmorphic erythrocytes (G1DE) or G1 cells in the detection of renal glomerular hematuria. Freshly voided urine samples from 174 patients with glomerular diseases were processed according to the method used for semiquantitative cytologic urinalysis. G1DEs (distorted erythrocytes with doughnut-like shape, target configuration with or without membranous protrusions or blebs), non-G1DEs (distorted erythrocytes without the above-mentioned morphologic changes), normal erythrocytes (NEs), and renal tubular cells (RTCs) were evaluated. Erythrocytic casts (ECs) were counted and graded as abundant (>1 per high-power field) or rare (1 per 5 high-power fields). G1DE/total erythrocyte ratios were calculated by counting 200 erythrocytes including G1DEs, non-G1DEs, and NEs. Only abundant NEs were found in 13 cases; abundant G1DEs, non-G1DEs, NEs, and no ECs in 95 cases; abundant NEs, non-G1DEs, and ECs and no G1DEs in 31 cases; and abundant NEs, G1DEs and non-G1DEs, and rare ECs in 35 cases. In 130 cases in which G1DEs were present, the G1DE/total erythrocyte ratios varied from 10% to 100%. This parameter was greater or equal to 80%, 50%, 20%, and 10% in 58 (44.6%), 29 (22.3%), 28 (21.5%), and 15 (11.5%) patients, respectively. In all cases, the number of RTCs was within normal limits or slightly increased, and a variable number of non-G1DEs were present in 161 cases. Thus, abundant ECs and/or G1DEs with a G1DE/total erythrocyte ratio of 10-100% proved to be specific urinary markers for renal glomerular diseases.


Pedunculated-type T1 colorectal carcinoma with lung carcinoma metastasis at the deepest invasive portion.
We present a rare case of colorectal T1 carcinoma with metastasis of previous lung carcinoma found at the deepest invasive portion. A 61-year-old man presented with cervical lymphadenopathy 18 years after undergoing surgery for right lung carcinoma [poorly differentiated adenocarcinoma stage IIb (T3N0M0)]. Contrast-enhanced computed tomography showed enlarged lymph nodes (LNs) in the neck and mediastinal regions. Combined hybrid-F-fluorodeoxyglucose positron emission-computerized tomography showed increased radionuclide uptake in multiple cervical LNs and mediastinal LNs. LN biopsy revealed a poorly differentiated adenocarcinoma, suspected to be a metastatic tumor of the lung. Subsequent colonoscopy revealed a pedunculated-type lesion with a depressed area in the ascending colon. We performed polypectomy as total excisional biopsy; this tumor was composed mainly of moderately differentiated adenocarcinoma, partially mixed with mucinous adenocarcinoma. The pathological findings of the invasive front of the colorectal carcinoma showed poorly differentiated adenocarcinoma with a morphological pattern similar to that of the previous lung carcinoma. Furthermore, immunohistochemical results for the histological type of the deepest invasive portion of the tissue specimen were positive for thyroid transcription factor-1 but negative for Caudal-type homeobox 2. From these morphological and immunohistochemical findings, the final diagnosis was moderately differentiated lung carcinoma, pTX N3 M1b (LN, colon) Stage IV.


Call to action: Prioritizing sleep health among US children and youth residing in alternative care settings.
Sleep health is a critical but under-recognized area of concern for the more than 650,000 children served by the US child welfare system each year. While sleep is vital to optimal child health and development, it is likely harmed by the multiple adversities and traumas experienced among children and youth residing in alternative care settings (ie, kinship care, nonrelative foster care, group homes). Children residing in alternative care settings have experienced, at a minimum, the trauma of removal from a biological parent's care and would benefit from holistic, comprehensive care approaches inclusive of sleep health. Furthermore, few studies are currently available to guide practitioners and policymakers in promoting sleep health among these children. In this Call to Action, our goal is to draw attention to the sleep health of children residing in alternative care settings. We highlight the need for a more robust evidence base to address major knowledge gaps and outline concrete steps toward building future promising sleep health-promoting practices and policies supporting children residing in alternative care settings.


[Clinicopathological implications of single positive core prostate cancer in a 12-core transrectal biopsy scheme].
To explore the clinicopathological implications of single positive core prostate cancer. A total of 45 patients with transrectal ultrasound (TRUS)-guided prostate biopsy were diagnosed with single positive core prostate cancer and subsequently underwent radical prostatectomy (RP). Their clinicopathological parameters were examined to identify the factors for disease upgrading or upstaging. On final pathology, only 31.1% patients had unilateral prostate cancer. And 37.8% patients experienced Gleason score upgrading, 66.7% patients had upstaging and 31.1% were found positive surgical margins. On multivariable analyses, the percentage of positive cores was an independent predictor of stage upgrading and perineural invasion at RP pathology. Age, prostate volume and preoperative prostate-specific antigen/prostate-specific antigen density (PSA/PSAD) had no significant effect on underestimated tumor burden. Most single positive core prostate cancer diagnosed by 12-core biopsy may be underestimated. The percentage of positive cores is an independent predictor of underestimated prostate cancer. Currently we have no instruments for accurately identifying microfocal or unilateral prostate cancer.


Adequacy and timeliness of antenatal care visits among Ethiopian women: a community-based panel study.
The purposes of the study are; (A) to identify whether Ethiopian women's antenatal care (ANC) visits are adequate and timely and (B) to explore factors that determine these. Panel study design. Ethiopia. A total of 2855 women nested within 217 enumeration areas. Adequacy and timeliness of ANC visits. Of all the 2855 respondents, 65% had made an ANC visit once, while 26.8% initiated ANC visits in a timely way and 43.3% attended adequate ANC visits. Rural residence (adjusted OR (AOR)=0.55, 95% CI: 0.36 to 0.84), attending higher level of education (AOR=2.64, 95% CI: 1.47 to 4.77), being multipara (AOR=0.53, 95% CI: 0.32 to 0.89) and encouragement by partners to attend clinic for ANC (AOR=1.98, 95% CI: 1.14 to 3.44) were significantly associated with timeliness of ANC visit. Similarly, residing in rural areas (AOR=0.20, 95% CI: 0.12 to 0.35), attending higher level of education (AOR=2.96, 95% CI: 1.38 to 6.15), encouragement by partners to attend clinic for ANC (AOR=2.11, 95% CI: 1.31 to 3.40) and timeliness of ANC visit (AOR=4.59, 95% CI: 2.93 to 7.21) were significantly associated with adequacy of ANC visits. A quarter of the pregnant women started ANC visits during the first trimester and nearly half attended adequate ANC visits with wider disparities across regions of their origin and their background characteristics. Concerted efforts on tailored interventions for rural residents, female education and partner involvement are recommended for early and adequate ANC visit(s).


Kikuchi's disease: a rare cause of cervical lymphadenitis and fever.
Cervical lymphadenitis and fever are common in patients presenting to the Emergency Department (ED). Kikuchi's disease is a rare, self-limited cause of fever and cervical lymphadenitis often misdiagnosed as lymphoma or lupus and inappropriately treated, potentially causing numerous ED visits for unrelieved symptoms. The case described is that of a 29-year-old with persistent fever and cervical lymphadenitis who presented to the ED with a suspected allergic reaction to an antibiotic. The diagnosis of Kikuchi's disease was made in association with nasopharyngeal carcinoma and partial hydatidiform mole. The case highlights the clinical features, diagnosis, and treatment of Kikuchi's disease.


Elevated levels of serum prolactin in patients with advanced multiple myeloma.
The role of prolactin in immunoregulation and normal hemopoiesis is well known. However, prolactin also seems to be involved in the pathomechanism of malignancies and autoimmune diseases. Elevated serum prolactin levels were reported in patients with malignant lymphoma, colon and breast carcinoma, systemic lupus erythematosus and rheumatoid arthritis. Recently we demonstrated prolactin immunostaining in bone marrow cells of patients with multiple myeloma. Serum prolactin levels of 56 patients with multiple myeloma, as well as serum beta(2)-microglobulin, and interleukin-6 concentrations were determined in this study. Patients with advanced disease showed a significant increase in serum prolactin concentration, while patients with a clinical stage of I and II, and also control patients had normal values. The concentration of serum beta(2)-microglobulin and interleukin-6 changed in parallel with that of serum prolactin in patients with multiple myeloma. Determining serum prolactin levels several times during the disease process in a given patient clearly showed that the prolactin concentration was increasing during the disease progression. Our results indicate a role of prolactin in disease progression in multiple myeloma.


On the pathogenesis of gastroesophageal reflux: the concept of gastroesophageal dyssynergia.
The cause of lower esophageal sphincter incompetence in gastroesophageal reflux disease is not clearly understood. We investigated the hypothesis that the esophagogastric junction incompetence results from failure of the gastric distention to produce the lower esophageal sphincter and crural diaphragm contraction caused by a disordered reflex action. The study was performed in 19 subjects (mean age, 42.6 +/- 7.2 years; 11 men and 8 women) who had reflux esophagitis and hiatus hernia and were scheduled for a fundoplication operation. Eight control volunteers (mean age, 41.8 +/- 6.9; 5 men and 3 women) who had huge supraumbilical ventral hernia but no reflux esophagitis or hiatus hernia were studied during operative hernia repair. The electromyographic activity and pressure response of the lower esophageal sphincter and crural diaphragm to separate esophageal and gastric distention were recorded. In the control subjects (volunteers) esophageal distention caused diminished electromyographic activity of the crural diaphragm and lower esophageal sphincter with decreased esophagogastric junction pressure, whereas gastric distention increased the electromyographic activity of the crural diaphragm and lower esophageal sphincter with increased esophagogastric junction pressure. In the patients the crural diaphragm and lower esophageal sphincter showed diminished resting electromyographic activity, with either no response or a paradoxical response to esophageal or gastric distention. The current study has demonstrated that the lower esophageal sphincter and crural diaphragm in patients with gastroesophageal reflux disease exhibited a diminished resting electric activity and either did not respond or reacted paradoxically to esophageal and gastric distention, constituting what we call esophagosphincteric and gastroesophageal paradox or dyssynergia. The cause of lower esophageal sphincter and crural diaphragm dysfunction is not known; a neurogenic cause was proposed. Further studies are required to investigate this point.


Scanning electron microscopy of crush/avulsion arterial trauma:effect of heparin and aspirin administration.
A model of vascular trauma and subsequent reanastomosis with poor postoperative likelihood of patency was developed in the rat femoral artery. Patency rates were significantly improved with intravenous heparin, intragastric aspirin, and both agents together. Heparin yielded higher patency than aspirin. Intimal surfaces of the vessels at various postoperative intervals were observed with the scanning electron microscope. More fibrin accumulation was seen in the aspirin-treated animals, while more platelet aggregation was found in the heparin-treated group. Both platelet aggregation and fibrin strand development appeared retarded with both agents administered. All groups demonstrated good healing of the ruptured intimal surfaces, beginning at 2 days postoperatively. It is concluded that fibrin strand development is a more significant factor in microvascular occlusion than platelet aggregation.


The prognosis of HPV-associated metastatic pharyngeal patients by primary and distant site.
Human papillomavirus (HPV) positivity is a favorable prognostic factor in the general population of head and neck squamous cell carcinoma (HNSCC) patients. However, its impact on the survival of metastatic HNSCC of pharynx (mHNSC-P) patients is unclear. This study aims to investigate the associations between HPV status and survival in mHNSC-P patients. 735 mHNSC-P patients diagnosed at first presentation from 2010 to 2016 were retrieved from the Surveillance, Epidemiology and End Result database (SEER). Chi-Squared test, univariate and multivariate cox proportional hazards model, Kaplan-Meier analysis, and log-rank test were applied to compare HPV-positive and -negative mHNSC-P patients. Using univariate cox proportional hazards analysis, HPV status, primary site, T stage, treatment and distant metastatic site correlate with the overall survival (OS) and disease-specific survival (DSS) in mHNSC-P patients. Multivariate cox regression analysis shows that HPV-positive mHNSC-P patients experienced significantly better OS (HR: 0.62 CI: 0.51-0.76, p < 0.001) and DSS (HR: 0.73 CI: 0.58-0.91, p < 0.01) as compared to HPV-negative mHNSC-P patients. Subgroup analysis indicates that HPV-associated OS and DSS benefits exist in patients with metastatic HNSCC of oropharynx (mHNSC-OP) but not in patients with metastatic HNSCC of non-oropharynx (mHNSC-non-OP). Among mHNSC-OP patients, HPV positivity confers disease-specific survival benefit in patients with oligometastatic rather than polymetastatic patients. Furthermore, HPV associated OS and DSS advantages in mHNSC-OP with lung metastasis was observed. HPV-positive mHNSC-OP patients with lung metastasis show better survival than HPV-negative mHNSC-OP patients, providing key information to guide patient treatment approaches.