Sporogonic development of a malaria parasite in vitro.
The sporogonic cycle of the avian malaria parasite Plasmodium gallinaceum was completed in vitro. Ookinetes (motile zygotes) were seeded onto a murine basement membrane-like gel (Matrigel) in coculture with Drosophila melanogaster cells (Schneider's L2). Transformation into oocysts as well as subsequent growth and differentiation were observed in parasites attached to Matrigel and depended on the presence of L2 cells. Sporozoites were first observed on day 10 in culture. Specific circumsporozoite protein antigenicity was identified in mature oocysts and in sporozoites. It is now possible to follow the entire life cycle of Plasmodium in vitro.


Novel INF2 mutations in an Italian cohort of patients with focal segmental glomerulosclerosis, renal failure and Charcot-Marie-Tooth neuropathy.
Mutations of INF2 represent the major cause of familial autosomal dominant (AD) focal segmental glomerulosclerosis (FSGS). A few patients present neurological symptoms of Charcot-Marie-Tooth (CMT) disease but the prevalence of the association has not been assessed yet. We screened 28 families with AD FSGS and identified 8 INF2 mutations in 9 families (32 patients overall), 3 of which were new. Mutations were in all cases localized in the diaphanous-inhibitory domain (DID) of the protein. Clinical features associated with INF2 mutations in our patient cohort included mild proteinuria (1.55 g/L; range 1-2.5) and haematuria as a unique symptom that was recognized at a median age of 21.75 years (range 8-30). Eighteen patients developed end-stage renal disease during their third decade of life; 12 patients presented a creatinine range between 1.2 and 1.5 mg/dL and 2 were healthy at 45 and 54 years of age. CMT was diagnosed in four cases (12.5%); one of these patients presented an already known mutation on exon 2 of INF2, whereas the other patients presented the same mutation on exon 4, a region that was not previously associated with CMT. We confirmed the high incidence of INF2 mutations in families with AD FSGS. The clinical phenotype was mild at the onset of the disease, but evolution to ESRD was frequent. The incidence of CMT has, for the first time, been calculated here to be 12.5% of mutation carriers. Our findings support INF2 gene analysis in families in which renal failure and/or neuro-sensorial defects are inherited following an AD model.


Reflux of Endoplasmic Reticulum proteins to the cytosol inactivates tumor suppressors.
In the past decades, many studies reported the presence of endoplasmic reticulum (ER)-resident proteins in the cytosol. However, the mechanisms by which these proteins relocate and whether they exert cytosolic functions remain unknown. We find that a subset of ER luminal proteins accumulates in the cytosol of glioblastoma cells isolated from mouse and human tumors. In cultured cells, ER protein reflux to the cytosol occurs upon ER proteostasis perturbation. Using the ER luminal protein anterior gradient 2 (AGR2) as a proof of concept, we tested whether the refluxed proteins gain new functions in the cytosol. We find that refluxed, cytosolic AGR2 binds and inhibits the tumor suppressor p53. These data suggest that ER reflux constitutes an ER surveillance mechanism to relieve the ER from its contents upon stress, providing a selective advantage to tumor cells through gain-of-cytosolic functions-a phenomenon we name ER to Cytosol Signaling (ERCYS).


MALDI/post ionization-ion mobility mass spectrometry of noncovalent complexes of dopamine receptors' epitopes.
Protein domains involved in receptor heteromer formation are disordered and rich in the amino acids necessary for the formation of noncovalent complexes (NCX). We present mass spectral NCX data from proteins and protein receptors' epitopes obtained by combining ion mobility (IM) and MALDI. We focus on NCX involved in heteromer formation occurring between epitopes of the Dopamine D2 (D2R) and Adenosine A2A receptors (A2AR) as well as D2R and the α2 nicotinic (NR) receptor's subunit. The IM data yield information on the gas phase conformation of the singly charged NCX which are observed either directly from MALDI or as codesorbed neutrals that are subsequently postionized by a time-delayed excimer laser pulse directed onto a portion of the neutral plume created by the MALDI desorption laser. Imaging mass spectrometry of the matrix/epitope dried droplet surface shows that the acidic and basic epitopes and their NCX are found to be spatially collocated within regions as small as 25 × 50 μm(2). Subtle differences in the relative abundance of protonated and cationized NCX and epitopes are measured in spatial regions near the sodium-rich outer border of the droplet.


Isolation and characterization of two alpha1-protease inhibitors in rat serum.
Two alpha1-protease inhibitors were identified in rat serum. On polyacrylamide gels, one (R-I) appeared at the leading edge of the albumin, the other (R-2) at the trailing edge. The two inhibitors have differing molecular weights, differing inhibitory spectra toward proteases, and are immunologically distinct. The assays for antiprotease activity were performed on whole human and rat sera and inhibitor-enriched fractions of rat serum that had been electrophoresed on polyacrylamide gels. The more rapidly migrating antiprotease of rat serum, R-I, inhibited trypsin, chymotrypsin, and elastase. The more slowly migrating antiprotease, R-2, inhibited both trypsin and chymotrypsin but possessed only weak anti-elastase activity. When a comparison was made between human alpha1-antitrypsin and the more rapidly migrating rat inhibitor, it could be seen that they have similar molecular weights, similar inhibitory spectra, and similar electrophoretic mobilities. The relevance of the study of the R-I inhibitor in the pathogenesis of experimental emphysema is emphasized.


Harnessing the Vnn1 pantetheinase pathway boosts short chain fatty acids production and mucosal protection in colitis.
In the management of patients with IBD, there is a need to identify prognostic markers and druggable biological pathways to improve mucosal repair and probe the efficacy of tumour necrosis factor alpha biologics. Vnn1 is a pantetheinase that degrades pantetheine to pantothenate (vitamin B5, a precursor of coenzyme A (CoA) biosynthesis) and cysteamine. Vnn1 is overexpressed by inflamed colonocytes. We investigated its contribution to the tolerance of the intestinal mucosa to colitis-induced injury. We performed an RNA sequencing study on colon biopsy samples from patients with IBD stratified according to clinical severity and modalities of treatment. We generated the VIVA mouse transgenic model, which specifically overexpresses Vnn1 on intestinal epithelial cells and explored its susceptibility to colitis. We developed a pharmacological mimicry of Vnn1 overexpression by administration of Vnn1 derivatives. VNN1 overexpression on colonocytes correlates with IBD severity. VIVA mice are resistant to experimentally induced colitis. The pantetheinase activity of Vnn1 is cytoprotective in colon: it enhances CoA regeneration and metabolic adaptation of colonocytes; it favours microbiota-dependent production of short chain fatty acids and mostly butyrate, shown to regulate mucosal energetics and to be reduced in patients with IBD. This prohealing phenotype is recapitulated by treating control mice with the substrate (pantethine) or the products of pantetheinase activity prior to induction of colitis. In severe IBD, the protection conferred by the high induction of VNN1 might be compromised because its enzymatic activity may be limited by lack of available substrates. In addition, we identify the elevation of indoxyl sulfate in urine as a biomarker of Vnn1 overexpression, also detected in patients with IBD. The induction of Vnn1/VNN1 during colitis in mouse and human is a compensatory mechanism to reinforce the mucosal barrier. Therefore, enhancement of vitamin B5-driven metabolism should improve mucosal healing and might increase the efficacy of anti-inflammatory therapy.


Pharmacological treatment for chronic oedema.
Chronic oedema is mainly treated by physical methods (compression, lymphatic drainage, massage and exercise), with drugs not having a major role to play. However, antibiotics are essential in the management of cellulitis, a common and important complication of chronic oedema. Diuretics and cortico-steroids may be appropriate in some types of oedema such as that associated with advanced cancer. Analgesics may be helpful in managing pain, particularly in the short term e.g. pain from cellulitis, although long-term use may also be appropriate, e.g. in cancer pain. Benzopyrones are not routinely used in the management of lymphoedema in the UK.


On the numerical evaluation of cuspoid diffraction catastrophes.
A simple computational approach is proposed for the evaluation of cuspoid diffraction catastrophes, which are fundamental tools for the description of wave fields in proximity to stable caustics. In particular, on representing the Pearcey and the swallowtail functions in terms of convergent series expansions, the action of the Weniger transformation on them is numerically studied for both real and complex values of the cuspoid arguments. Numerical experiments, aimed at giving evidence of the effectiveness and implementative ease of the proposed method, together with some quantitative comparisons to previously published results, are also presented.


Knowledge, Attitude, and Practice of Obstetric Nurses in Relation to Breast Cancer and Breast Self-examination.
This study aims to determine obstetrics nurses knowledge, attitude, and practice about breast cancer and breast self-examination (BSE), and to contribute to the early detection of breast cancer during breastfeeding periods. For the survey, 163 individuals (obstetric nurses) were chosen by convenience sampling and agreed to participate in the study. Their knowledge, attitude, and practice about breast cancer and BSE. Knowledge and attitude averaged 70.45+/-10.90 (of 100) and 3.64+/-0.27 (of 5), respectively. While most nurses (99%) recognized the importance of BSE, only 58.9% experienced BSE. BSE practice level averaged 8.35+/-1.96 (of 12). Only 20.2% had recommended BSE to their clients. Practice level varied significantly for different marital status, breastfeeding experience, and education, while knowledge and attitude remained independent. Nurses who had experienced mammogram or breast ultrasonogram themselves scored higher in knowledge. Attitude was higher for nurses who received recommendation for BSE, performed BSE, received BSE education, or recommended BSE to clients. Practice level was higher for nurses who received BSE education or willing to perform BSE in future. Practice level had a positive correlation with attitude but no correlation to knowledge. Obstetric nurses need continuing education for practicing BSE. Practical BSE education can not only promote preventive behavior of nurses, but it can also improve the breast health management of obstetrical clients.


DriverMP enables improved identification of cancer driver genes.
Cancer is widely regarded as a complex disease primarily driven by genetic mutations. A critical concern and significant obstacle lies in discerning driver genes amid an extensive array of passenger genes. We present a new method termed DriverMP for effectively prioritizing altered genes on a cancer-type level by considering mutated gene pairs. It is designed to first apply nonsilent somatic mutation data, protein‒protein interaction network data, and differential gene expression data to prioritize mutated gene pairs, and then individual mutated genes are prioritized based on prioritized mutated gene pairs. Application of this method in 10 cancer datasets from The Cancer Genome Atlas demonstrated its great improvements over all the compared state-of-the-art methods in identifying known driver genes. Then, a comprehensive analysis demonstrated the reliability of the novel driver genes that are strongly supported by clinical experiments, disease enrichment, or biological pathway analysis. The new method, DriverMP, which is able to identify driver genes by effectively integrating the advantages of multiple kinds of cancer data, is available at https://github.com/LiuYangyangSDU/DriverMP. In addition, we have developed a novel driver gene database for 10 cancer types and an online service that can be freely accessed without registration for users. The DriverMP method, the database of novel drivers, and the user-friendly online server are expected to contribute to new diagnostic and therapeutic opportunities for cancers.


Radiologist-rheumatologist multidisciplinary approach in the management of axial spondyloarthritis: a Delphi consensus statement.
The aim of this study was to develop a Delphi consensus statement between rheumatologists and radiologists for the diagnosis and monitoring of axial spondyloarthritis (axial-SpA). Following an extensive literature search to identify unmet needs and potential goals for a multidisciplinary approach, a scientific board comprising 28 Italian hospital-based rheumatologists (n=19) and radiologists (n=9) identified 8 "starting points", resulting in the development of 23 consensus statements covering issues from current practice guidelines to specific MRI protocols for the assessment of axial-SpA. Each participant anonymously expressed a level of agreement for each statement using a 5-point Likert scale (1="strongly disagree"; 5="strongly agree") via an online Delphi method.Total cumulative agreement (TCA) was defined as the sum of the percentage of response to items 4 ("agree") and 5 ("absolutely agree"). Consensus was defined as ≥80% total cumulative agreement for each statement. After the first round of voting (28 participants), positive consensus was reached for 28/31 (90.3%) statements. Statements without consensus (n=3) were discussed in a face-to-face plenary session prior to the second vote (28 participants). After the second round voting, positive consensus was attained for all 31 statements, with mean final TCA of 95.5% (range 82.1-100%). This Delphi consensus statement provides an aid to rheumatologists and radiologists for the diagnosis and monitoring of axial-SpA.


Spontaneous regression of pulmonary metastases from testicular embryonal carcinoma.
A 22-year-old man was referred to our hospital for a lung tumor on a chest X-ray examination. Computed tomography of the chest revealed multiple coin lesions. Magnetic resonance imaging of the left testis showed an 8-mm tumor that was hard and palpable. Testicular tumor with lung metastasis was diagnosed and orchiectomy was performed. The histopathological diagnosis was embryonal carcinoma with infiltration of histiocytes. Four days after the operation, a chest X-ray showed a remarkable regression of the lung tumors. Chemotherapy was not performed because the metastatic lesions continued to regress spontaneously. Six months later, no tumor was observed on computed tomography images of the lungs.


[Disabling pathogenetic causes in postthrombotic disease of the lower extremities].
Prospective investigation of etiological and pathogenetic causes of the disabling complications incidence in the lower extremities postthrombotic disease (LEPTHD), influencing activity of these patients, was conducted. The examined patients were divided into two groups, in 62 (58.5%) patients a disability was absent, and in 44 (41.5%) disability was established. Profound clinical examination was conducted, including determination of subfascial pressure on the shin, ultrasound duplex scanning of venous system, electroneuromyography of the lower extremities, estimation of the D-dimer, levels antithrombine-III activity in general and regional blood flow. The leading factors, which causes the LEPTHD patients activity restriction, were determined, basing on the results analysis.


[Effect of unithiol on the ultrastructure of the myocardium and adrenals in endotoxin shock].
Administration of unithiol to animals with endotoxin shock promotes deposition of catecholamines in chromaffin cells of the adrenals, blocks their secretion to the bloodstream and prevents the development of injuries to the cortical substance of the gland. Administration of unithiol for endotoxemia prevents damage to intracellular organelles of cardiomyocytes. However, insignificant disorders of the contractile apparatus of the cells and histohematic barriers are maintained. The capacity of unithiol to prevent the progress of destructive alterations in the heart and adrenals permits recommending it for use in combined treatment of septic conditions.


Breast cancer risks and prevention: implications of the Breast Cancer Prevention Trial results.
Breast cancer is a major health concern for women in the U.S. Several factors determine a woman's risk for developing breast cancer including age, genetics, hormonal exposure and others. It is possible to estimate a woman's risk for developing breast cancer by using a statistical model known as the Gail model. Using this model, the Breast Cancer Prevention Trial (BCPT) found that women at high risk for breast cancer could have their incidence of breast cancer reduced by 49% by taking tamoxifen. The serious side effects of tamoxifen therapy were almost exclusively seen in women over age 50. Women under 50 with a high risk of breast cancer are, therefore, most likely to benefit from taking tamoxifen for five years. Thus, a benefit-to-risk estimate should be made for each woman before advising her to take tamoxifen. A new trial (Study of Tamoxifen And Raloxifene, STAR) will compare tamoxifen with raloxifene for their effectiveness in reducing breast cancer and for side effects. West Virginia women participated in the BCPT and will be able to participate in the STAR trial through centers in Morgantown and Charleston.


The effect of pasture quantity temporal variation on milking robot utilization.
In pasture-based automatic milking systems (AMS), a decrease in robot utilization (RU) often occurs in the early morning hours. Novel feeding strategies that encourage voluntary cow traffic throughout 24 h could help mitigate this problem. We determined the effect of 3 distinct pasture allocation methods on RU patterns throughout a 24-h period. The experiment was conducted at the University of Melbourne's Dookie research farm in northern Victoria, Australia. Three Lely Astronaut A3 robotic milking units (Lely, Maassluis, the Netherlands) milked 133 cows, grazing pasture, with concentrate offered at milking in the robots. The farm operated a system of 3-way grazing, with active access to each pasture allocation: 2030-0400 h (allocation A), 0400-1330 h (allocation B), and 1330-2030 h (allocation C). Treatments varied in the quantity of feed offered per hour of active access to each of the 3 pasture allocations. The control treatment offered the same proportion of feed (corrected for active access time) in all 3 pasture allocations (allocation A = 31.3%, B = 39.6%, and C = 29.2%). The day treatment offered the largest proportion of feed during the day (allocation A = 20%, B = 40%, and C = 40%), following the cows' diurnal pattern of feeding activity. The night treatment offered the largest proportion of feed at night (allocation A = 42%, B = 40%, and C = 18%). Due to the nature of pasture-based AMS, treatments could not be applied simultaneously. Therefore, treatments were applied to the entire herd and repeated twice over 42 d, lasting 7 d/treatment, with the first 3 d for habituation, followed by 4 d of data collection. Robot utilization (milkings/h) varied throughout 24 h between treatments, with the night treatment recording greater RU at 0800, 1800, and 1900 h and lower RU between 2100 to 0100 h, compared with the day treatment. The proportion of the herd milking between 0000 and 0600 h was greater for the control (43.3%) and day (45.3%) treatments compared with the night treatment (25.8%). Herd-average daily pasture intake was similar (10.5 kg of dry matter) for all treatments. This experiment is the first to demonstrate the manipulation of RU by varying the quantity of pasture offered. However, the use of variable allocation alone did not eliminate the decrease in RU between 0000 and 0600 h, with the timing of allocation also likely to play a role. We recommend a further research focus on combining both timing and quantity of pasture allocated to improve RU in pasture-based AMS.


Contact and encirclement of glioma cells in vitro is an intrinsic behavior of a clonal human neural stem cell line.
Pathotropic neural stem and/or progenitor cells (NSCs) can potentially deliver therapeutic agents to otherwise inaccessible cancers. In glioma, NSCs are found in close contact with tumor cells, raising the possibility that specificity of NSC contact with glioma targets originates in the tumor cells themselves. Alternatively, target preferences may originate, at least in part, in the tumor microenvironment. To better understand mechanisms underlying NSC interactions with glioma cells, we examined NSC-target cell contacts in a highly simplified 3-dimensional peptide hydrogel (Puramatrix) in which cell behaviors can be studied in the relative absence of external cues. HB1.F3 is an immortalized clonal human NSC line extensively characterized in preclinical investigations. To study contact formation between HB1.F3 NSCs and glioma cells, we first examined co-cultures of eGFP-expressing HB1.F3 (HB1.F3.eGFP) NSCs and dsRed-expressing U251 glioma (U251.dsRed) cells. Using confocal microscopy, HB1.F3.eGFP cells were observed contacting or encircling U251.dsRed glioma cells, but never the reverse. Next, examining specificity of these contacts, no significant quantitative differences in either percentages of HB1.F3 NSCs contacting targets, or in the extent of target cell encirclement, were observed when HB1.F3.eGFP cells were presented with various potential target cells (human glioma and breast cancer cell lines, patient-derived brain tumor lines, non-tumor fibroblasts, primary mouse and human astroglial cells, and primary adult and newborn human dermal fibroblasts) except that interactions between HB1.F3 cells did not progress beyond establishing contacts. Finally cytoskeletal mechanisms employed by HB1.F3.eGFP cells varied with the substrate. When migrating in Puramatrix, HB1.F3 NSCs exhibited intermittent process extension followed by soma translocation, while during encirclement their movements were more amoeboid. We conclude that formation of contacts and subsequent encirclement of target cells by HB1.F3 NSCs is an intrinsic property of these NSCs, and that preferential contact formation with tumor cells in vivo must therefore be highly dependent on microenvironmental cues.


Acute ankle injuries: clinical decision rules for radiographse.
The need to perform "routine" radiographs after every case of ankle trauma has been repeatedly questioned, since less than 15 percent of ankle injuries are found to involve a significant fracture. Several authors have proposed guidelines to define clinical characteristics that may help physicians identify patients with a higher probability of having a fracture on the radiograph. The Ottawa ankle rules are the latest guidelines developed for the management of ankle injuries. These highly sensitive decision rules may allow a significant reduction in the number of ankle radiographic series ordered and may decrease patients' waiting times and costs without an increased rate of missed fractures or patient dissatisfaction.


IL-12 cDNA direct injection: antimetastatic effect from a single injection in a murine hepatic metastases model.
Interleukin 12 (IL-12) gene therapy is an effective antitumor agent in local and metastatic murine tumor models. We sought to evaluate the antimetastatic effect of IL-12 cDNA in a liver metastases model. A liver metastases model was induced by creating a "primary" splenic tumor through inoculation of 1 x 10(5) TS/A adenocarcinoma cells directly into the inferior pole of the spleen in female BALB/c mice. On day 4, 50 microg of IL-12 cDNA or control plasmid DNA was injected into splenic tumor, followed by splenectomy on day 8. Mice were sacrificed on day 25 to assess liver tumor burden. IL-12 mRNA and mIL-12 and IFN-gamma protein levels were assessed after IL-12 injection. Peripheral blood CD4+, CD8+, and NK cells were quantified on day 14 using FACS. To determine the significance of site of cytokine DNA injection, IL-12 cDNA was injected on day 4 into splenic tumor or into the non-involved spleen after isolation of the inferior and superior portions of the spleen, respectively, with surgical clips. Splenectomy was performed on day 8 and sacrifice was performed on day 25. IL-12 mRNA was detected in the liver 8 h after injection, with a peak at 24 h. After splenic injection, protein levels of IL-12 and IFN-gamma were detectable in the liver and spleen 24 h after treatment. IL-12 and IFN-gamma were not detectable in control animals. In the peripheral blood, there was a marked increase in NK cells (13% of total lymphocytes versus 4%, control) and in the CD4+/CD8+ ratio (5.5 versus 1.9). At day 25, there was a marked antimetastatic effect after IL-12 injection into either splenic tumor [liver:body weight, 6.2 versus 10.9 (control), P = 0.007] or non-involved spleen (6.8 g versus 10.7 g, P = 0.005). There was no difference in the antimetastatic effect between animals injected into splenic tumor or non-involved spleen (P = 0.3). Injection with a single dose of IL-12 cDNA into splenic tumor or non-involved spleen resulted in a profound antimetastatic effect. Splenic IL-12 injection results in mRNA expression in the liver, protein expression in the liver and spleen, and a marked increase in NK cells and the CD4+/CD8+ ratio in peripheral blood.


Juvenile hormone (JH) esterase of the mosquito Culex quinquefasciatus is not a target of the JH analog insecticide methoprene.
Juvenile hormones (JHs) are essential sesquiterpenes that control insect development and reproduction. JH analog (JHA) insecticides such as methoprene are compounds that mimic the structure and/or biological activity of JH. In this study we obtained a full-length cDNA, cqjhe, from the southern house mosquito Culex quinquefasciatus that encodes CqJHE, an esterase that selectively metabolizes JH. Unlike other recombinant esterases that have been identified from dipteran insects, CqJHE hydrolyzed JH with specificity constant (k(cat)/K(M) ratio) and V(max) values that are common among JH esterases (JHEs). CqJHE showed picomolar sensitivity to OTFP, a JHE-selective inhibitor, but more than 1000-fold lower sensitivity to DFP, a general esterase inhibitor. To our surprise, CqJHE did not metabolize the isopropyl ester of methoprene even when 25 pmol of methoprene was incubated with an amount of CqJHE that was sufficient to hydrolyze 7,200 pmol of JH to JH acid under the same assay conditions. In competition assays in which both JH and methoprene were available to CqJHE, methoprene did not show any inhibitory effects on the JH hydrolysis rate even when methoprene was present in the assay at a 10-fold higher concentration relative to JH. Our findings indicated that JHE is not a molecular target of methoprene. Our findings also do not support the hypothesis that methoprene functions in part by inhibiting the action of JHE.


Effect of short dexamethasone suppression on plasma steroids in prepubertal and pubertal girls.
In 40 girls aged from 2 to 14 years, subdivided into groups according to age and pubertal development, and in 6 adult female volunteers, plasma cortisol (F), pregnenolone (delta 5), dehydroepiandrosterone (DHA) progesterone (P), 17-hydroxyprogesterone (17P), androstenedione (A), testosterone (T) and estradiol (E2) were measured before and after short dexamethasone (DXM) suppression. The results confirmed the capacity of DXM to inhibit plasma steroids in all age groups, except T in 2-9 year old and P1 Tanner's stage girls. The percentage suppression of each given steroid was constant over the age groups from 6-9 years to P4-5 Tanner's stage, while lower suppression was found in 17P, P and DHA in 2-5 year old girls and in 17P, DHA and E2 in adult women. These results emphasize the fundamental role of ACTH as the overall stimulating factor of adrenal steroidogenesis but do not negate the possibility of another factor responsible for the development of the adrenal androgen secreting cells throughout prepuberty and puberty.


The effects of quality of evidence communication on perception of public health information about COVID-19: Two randomised controlled trials.
The quality of evidence about the effectiveness of non-pharmaceutical health interventions is often low, but little is known about the effects of communicating indications of evidence quality to the public. In two blinded, randomised, controlled, online experiments, US participants (total n = 2140) were shown one of several versions of an infographic illustrating the effectiveness of eye protection in reducing COVID-19 transmission. Their trust in the information, understanding, feelings of effectiveness of eye protection, and the likelihood of them adopting it were measured. Compared to those given no quality cues, participants who were told the quality of the evidence on eye protection was 'low', rated the evidence less trustworthy (p = .001, d = 0.25), and rated it as subjectively less effective (p = .018, d = 0.19). The same effects emerged compared to those who were told the quality of the evidence was 'high', and in one of the two studies, those shown 'low' quality of evidence said they were less likely to use eye protection (p = .005, d = 0.18). Participants who were told the quality of the evidence was 'high' showed no statistically significant differences on these measures compared to those given no information about evidence quality. Without quality of evidence cues, participants responded to the evidence about the public health intervention as if it was high quality and this affected their subjective perceptions of its efficacy and trust in the provided information. This raises the ethical dilemma of weighing the importance of transparently stating when the evidence base is actually low quality against evidence that providing such information can decrease trust, perception of intervention efficacy, and likelihood of adopting it.


Dopaminergic regulation of the estrogen-induced glandular kallikrein in the rat anterior pituitary.
Glandular kallikrein is a major estrogen-induced protein of the rat anterior pituitary. A second kallikrein-like protease in the rat anterior pituitary (kallikrein A) is not affected by estrogens, nor is a third pituitary protease which cleaves a trypsin substrate but not kallikrein substrates. This study examined whether any of the pituitary proteases are regulated by dopaminergic mechanisms. Ovariectomized female rats were treated for 5-10 days with reserpine (a catecholamine depleting agent), haloperidol (a dopamine receptor blocker) or bromocriptine (a dopamine receptor agonist); some rats also received 1 or 2 micrograms estradiol benzoate every 48 h. Following activation of latent proteases with trypsin, anterior pituitary extracts were assayed for kallikrein activity before and after fractionation on DEAE-Sephadex to separate the two kallikrein-like proteases. Reserpine or haloperidol doubled glandular kallikrein levels in anterior pituitaries from estrogen-treated rats. Reserpine or haloperidol had little or no effect in the absence of estrogen (estrogen produced a 5- to 7-fold increase in glandular kallikrein in the absence of drug treatment). Bromocriptine markedly attenuated the ability of estrogen to induce glandular kallikrein. Further, bromocriptine blocked the ability of reserpine to increase glandular kallikrein levels, and haloperidol attenuated the effect of bromocriptine. Other anterior pituitary proteases were unaffected by either estrogen, haloperidol, reserpine or bromocriptine. The results demonstrate that the estrogen induction of glandular kallikrein in the rat anterior pituitary is modulated by inhibitory dopaminergic mechanisms. Prolactin is the only pituitary hormone which exhibits a similar profile of hormonal and neuroendocrine regulation; this suggests a possible link between glandular kallikrein and prolactin.


A model-based assessment of the cost-utility of strategies to identify Lynch syndrome in early-onset colorectal cancer patients.
Lynch syndrome is an autosomal dominant cancer predisposition syndrome caused by mutations in the DNA mismatch repair genes MLH1, MSH2, MSH6 and PMS2. Individuals with Lynch syndrome have an increased risk of colorectal cancer, endometrial cancer, ovarian and other cancers. Lynch syndrome remains underdiagnosed in the UK. Reflex testing for Lynch syndrome in early-onset colorectal cancer patients is proposed as a method to identify more families affected by Lynch syndrome and offer surveillance to reduce cancer risks, although cost-effectiveness is viewed as a barrier to implementation. The objective of this project was to estimate the cost-utility of strategies to identify Lynch syndrome in individuals with early-onset colorectal cancer in the NHS. A decision analytic model was developed which simulated diagnostic and long-term outcomes over a lifetime horizon for colorectal cancer patients with and without Lynch syndrome and for relatives of those patients. Nine diagnostic strategies were modelled which included microsatellite instability (MSI) testing, immunohistochemistry (IHC), BRAF mutation testing (methylation testing in a scenario analysis), diagnostic mutation testing and Amsterdam II criteria. Biennial colonoscopic surveillance was included for individuals diagnosed with Lynch syndrome and accepting surveillance. Prophylactic hysterectomy with bilateral salpingo-oophorectomy (H-BSO) was similarly included for women diagnosed with Lynch syndrome. Costs from NHS and Personal Social Services perspective and quality-adjusted life years (QALYs) were estimated and discounted at 3.5% per annum. All strategies included for the identification of Lynch syndrome were cost-effective versus no testing. The strategy with the greatest net health benefit was MSI followed by BRAF followed by diagnostic genetic testing, costing £5,491 per QALY gained over no testing. The effect of prophylactic H-BSO on health-related quality of life (HRQoL) is uncertain and could outweigh the health benefits of testing, resulting in overall QALY loss. Reflex testing for Lynch syndrome in early-onset colorectal cancer patients is predicted to be a cost-effective use of limited financial resources in England and Wales. Research is recommended into the cost-effectiveness of reflex testing for Lynch syndrome in other associated cancers and into the impact of prophylactic H-BSO on HRQoL.


Loss of ABCG1 results in chronic pulmonary inflammation.
ABCG1, a member of the ATP-binding cassette transporter superfamily, is highly expressed in multiple cells of the lung. Loss of ABCG1 results in severe pulmonary lipidosis in mice, with massive deposition of cholesterol in both alveolar macrophages and type 2 cells and the accumulation of excessive surfactant phospholipids. These observations are consistent with ABCG1 controlling cellular sterol metabolism. Herein, we report on the progressive and chronic inflammatory process that accompanies the lipidosis in the lungs of Abcg1-/- mice. Compared with wild-type animals, the lungs of aged chow-fed mice deficient in ABCG1 show distinctive signs of inflammation that include macrophage accumulation, lymphocytic infiltration, hemorrhage, eosinophilic crystals, and elevated levels of numerous cytokines and cytokine receptors. Analysis of bronchoalveolar lavages obtained from Abcg1-/- mice revealed elevated numbers of foamy macrophages and leukocytes and the presence of multiple markers of inflammation including crystals of chitinase-3-like proteins. These data suggest that cholesterol and/or cholesterol metabolites that accumulate in Abcg1-/- lungs can trigger inflammatory signaling pathways. Consistent with this hypothesis, the expression of a number of cytokines was found to be significantly increased following increased cholesterol delivery to either primary peritoneal macrophages or Raw264.7 cells. Finally, cholesterol loading of primary mouse macrophages induced cytokine mRNAs to higher levels in Abcg1-/-, as compared with wild-type cells. These results demonstrate that ABCG1 plays critical roles in pulmonary homeostasis, balancing both lipid/cholesterol metabolism and inflammatory responses.


Fine-scale genetic structuring in a natural population of European wild rabbits (Oryctolagus cuniculus).
The genetic structure of a free-living tagged population of European wild rabbits (Oryctolagus cuniculus) was investigated for two consecutive years (1990 and 1991) using 10 polymorphic microsatellite loci. A specific social behaviour, the formation of stable breeding groups, influenced the genetic structure of the population. These breeding groups were shown to constitute genetically differentiated units with low levels of gene flow between them. The average relatedness among members of a social group was higher than within the population as a whole. As a result of female philopatry coupled with male-biased natal dispersal, the relatedness of females was higher than that of males, both within social groups and in the whole population. Furthermore, the average relatedness of females within groups was twice the relatedness of females between groups. This study reveals marked fine-scale, intrapopulation genetic structure, which is attributable to the social behaviour of the European wild rabbit.


Enantioselective Synthesis of Vicinal (R,R)-Diols by Saccharomyces cerevisiae Butanediol Dehydrogenase.
Butanediol dehydrogenase (Bdh1p) from Saccharomyces cerevisiae belongs to the superfamily of the medium-chain dehydrogenases and reductases and converts reversibly R-acetoin and S-acetoin to (2R,3R)-2,3-butanediol and meso-2,3-butanediol, respectively. It is specific for NAD(H) as a coenzyme, and it is the main enzyme involved in the last metabolic step leading to (2R,3R)-2,3-butanediol in yeast. In this study, we have used the activity of Bdh1p in different forms-purified enzyme, yeast extracts, permeabilized yeast cells, and as a fusion protein (with yeast formate dehydrogenase, Fdh1p)-to transform several vicinal diketones to the corresponding diols. We have also developed a new variant of the delitto perfetto methodology to place BDH1 under the control of the GAL1 promoter, resulting in a yeast strain that overexpresses butanediol dehydrogenase and formate dehydrogenase activities in the presence of galactose and regenerates NADH in the presence of formate. While the use of purified Bdh1p allows the synthesis of enantiopure (2R,3R)-2,3-butanediol, (2R,3R)-2,3-pentanediol, (2R,3R)-2,3-hexanediol, and (3R,4R)-3,4-hexanediol, the use of the engineered strain (as an extract or as permeabilized cells) yields mixtures of the diols. The production of pure diol stereoisomers has also been achieved by means of a chimeric fusion protein combining Fdh1p and Bdh1p. Finally, we have determined the selectivity of Bdh1p toward the oxidation/reduction of the hydroxyl/ketone groups from (2R,3R)-2,3-pentanediol/2,3-pentanedione and (2R,3R)-2,3-hexanediol/2,3-hexanedione. In conclusion, Bdh1p is an enzyme with biotechnological interest that can be used to synthesize chiral building blocks. A scheme of the favored pathway with the corresponding intermediates is proposed for the Bdh1p reaction.


Analytic Energy Gradients for Variational Two-Electron Reduced-Density-Matrix-Driven Complete Active Space Self-Consistent Field Theory.
Analytic energy gradients are presented for a variational two-electron reduced-density-matrix (2-RDM)-driven complete active space self-consistent field (CASSCF) method. The active-space 2-RDM is determined using a semidefinite programing (SDP) algorithm built upon an augmented Lagrangian formalism. Expressions for analytic gradients are simplified by the fact that the Lagrangian is stationary with respect to variations in both the primal and the dual solutions to the SDP problem. Orbital response contributions to the gradient are identical to those that arise in conventional CASSCF methods in which the electronic structure of the active space is described by a full configuration interaction (CI) wave function. We explore the relative performance of variational 2-RDM (v2RDM)- and CI-driven CASSCF for the equilibrium geometries of 20 small molecules. When enforcing two-particle N-representability conditions, full-valence v2RDM-CASSCF-optimized bond lengths display a mean unsigned error of 0.0060 Å and a maximum unsigned error of 0.0265 Å, relative to those obtained from full-valence CI-CASSCF. When enforcing partial three-particle N-representability conditions, the mean and maximum unsigned errors are reduced to only 0.0006 and 0.0054 Å, respectively. For these same molecules, full-valence v2RDM-CASSCF bond lengths computed in the cc-pVQZ basis set deviate from experimentally determined ones on average by 0.017 and 0.011 Å when enforcing two- and three-particle conditions, respectively, whereas CI-CASSCF displays an average deviation of 0.010 Å. The v2RDM-CASSCF approach with two-particle conditions is also applied to the equilibrium geometry of pentacene; optimized bond lengths deviate from those derived from experiment, on average, by 0.015 Å when using a cc-pVDZ basis set and a (22e,22o) active space.


Live imaging of the cytoskeleton in early cleavage-stage zebrafish embryos.
The large and transparent cells of cleavage-stage zebrafish embryos provide unique opportunities to study cell division and cytoskeletal dynamics in very large animal cells. Here, we summarize recent progress, from our laboratories and others, on live imaging of the microtubule and actin cytoskeletons during zebrafish embryonic cleavage. First, we present simple protocols for extending the breeding competence of zebrafish mating ensembles throughout the day, which ensures a steady supply of embryos in early cleavage, and for mounting these embryos for imaging. Second, we describe a transgenic zebrafish line [Tg(bactin2:HsENSCONSIN17-282-3xEGFP)hm1] that expresses the green fluorescent protein (GFP)-labeled microtubule-binding part of ensconsin (EMTB-3GFP). We demonstrate that the microtubule-based structures of the early cell cycles can be imaged live, with single microtubule resolution and with high contrast, in this line. Microtubules are much more easily visualized using this tagged binding protein rather than directly labeled tubulin (injected Alexa-647-labeled tubulin), presumably due to lower background from probe molecules not attached to microtubules. Third, we illustrate live imaging of the actin cytoskeleton by injection of the actin-binding fragment of utrophin fused to GFP. Fourth, we compare epifluorescence-, spinning-disc-, laser-scanning-, and two-photon-microscopic modalities for live imaging of the microtubule cytoskeleton in early embryos of our EMTB-3GFP-expressing transgenic line. Finally, we discuss future applications and extensions of our methods.


Proteolysis-independent downregulation of DELLA repression in Arabidopsis by the gibberellin receptor GIBBERELLIN INSENSITIVE DWARF1.
This article presents evidence that DELLA repression of gibberellin (GA) signaling is relieved both by proteolysis-dependent and -independent pathways in Arabidopsis thaliana. DELLA proteins are negative regulators of GA responses, including seed germination, stem elongation, and fertility. GA stimulates GA responses by causing DELLA repressor degradation via the ubiquitin-proteasome pathway. DELLA degradation requires GA biosynthesis, three functionally redundant GA receptors GIBBERELLIN INSENSITIVE DWARF1 (GID1a, b, and c), and the SLEEPY1 (SLY1) F-box subunit of an SCF E3 ubiquitin ligase. The sly1 mutants accumulate more DELLA proteins but display less severe dwarf and germination phenotypes than the GA biosynthesis mutant ga1-3 or the gid1abc triple mutant. Interestingly, GID1 overexpression rescued the sly1 dwarf and infertility phenotypes without decreasing the accumulation of the DELLA protein REPRESSOR OF ga1-3. GID1 rescue of sly1 mutants was dependent on the level of GID1 protein, GA, and the presence of a functional DELLA motif. Since DELLA shows increasing interaction with GID1 with increasing GA levels, it appears that GA-bound GID1 can block DELLA repressor activity by direct protein-protein interaction with the DELLA domain. Thus, a SLY1-independent mechanism for GA signaling may function without DELLA degradation.


[Effect of succinylcholine on recovery of atracurium-induced neuromuscular blockade].
If muscle relaxation is inadequate at the end of surgery, a transient increase in neuromuscular (NM) blockade should be needed to facilitate closure. Succinylcholine is the only agent of short duration available currently and could be used in the just mentioned circumstance. Therefore, the study was designed to investigate the effects of succinylcholine on the recovery of atracurium-induced NM blockade. Sixty-four female patients who underwent elective surgery under general anesthesia were allocated into 6 groups. Before anesthesia, no patient had received drugs which affect NM transmission. In all patients anesthesia was induced with thiopental, diazepam and fentanyl and maintained with 50% nitrous oxide in oxygen and 0.5-1.5% isoflurane. During anesthesia, blood pressure and electrocardiogram were monitored and end-tidal CO2 was maintained at 30-40 mmHg. Control records of the integrated electromyography responded to train-of-four supramaximal stimulation using a Datex Relaxograph were obtained after induction of anesthesia before a block was produced with an initial dose of succinylcholine 1.0 mg in group I (n = 8) or atracurium 0.4 mg/kg in groups II-VI. On first twitch (T1) recovering to 50% of control, 0.4 mg/kg, 0.6 mg/kg, 0.8 mg/kg, 1.0 mg/kg and 1.2 mg/kg of succinylcholine were given in groups II (n = 11), III (n = 14), IV (n = 11), V (n = 10), VI (n = 10) respectively. It was seen that succinylcholine 0.4 mg/kg antagonized NM blockade by atracurium but at 0.6 mg/kg, 0.8 mg/kg, 1.0 mg/kg, 1.2 mg/kg it enhanced that.(ABSTRACT TRUNCATED AT 250 WORDS)


Metagenomics harvested genus-specific single-stranded DNA-annealing proteins improve and expand recombineering in Pseudomonas species.
The widespread Pseudomonas genus comprises a collection of related species with remarkable abilities to degrade plastics and polluted wastes and to produce a broad set of valuable compounds, ranging from bulk chemicals to pharmaceuticals. Pseudomonas possess characteristics of tolerance and stress resistance making them valuable hosts for industrial and environmental biotechnology. However, efficient and high-throughput genetic engineering tools have limited metabolic engineering efforts and applications. To improve their genome editing capabilities, we first employed a computational biology workflow to generate a genus-specific library of potential single-stranded DNA-annealing proteins (SSAPs). Assessment of the library was performed in different Pseudomonas using a high-throughput pooled recombinase screen followed by Oxford Nanopore NGS analysis. Among different active variants with variable levels of allelic replacement frequency (ARF), efficient SSAPs were found and characterized for mediating recombineering in the four tested species. New variants yielded higher ARFs than existing ones in Pseudomonas putida and Pseudomonas aeruginosa, and expanded the field of recombineering in Pseudomonas taiwanensisand Pseudomonas fluorescens. These findings will enhance the mutagenesis capabilities of these members of the Pseudomonas genus, increasing the possibilities for biotransformation and enhancing their potential for synthetic biology applications. .


Mll5 is required for normal spermatogenesis.
Mll5 is currently a member of the Mll family of SET domain histone methyltransferase proteins but studies have also showed that it could be part of the SET3 branch of proteins. Recently, constitutive knock out animal studies have shown that Mll5 is required for proper haematopoietic stem cell differentiation, and loss of Mll5 results in synthetic lethality for genome de-methylation. Mll5 deficient male mice are infertile and here we analyse the consequences of Mll5 deficiency for spermatogenesis. Mll5 deficient male mice, but not female mice, are infertile. Here we show using RNA in-situ hybridization that Mll5 is expressed in the germ cells of the testes of wild type mice. Consistent with the expression of Mll5, we demonstrate by electron microscopy, video microscopy and in vitro fertilisation techniques that Mll5 deficient mice have defects in terminal maturation and packaging of sperm. The defects seen include detachment of the acrosomal cap and impaired excess cytoplasm removal. Functional tests of sperm motility show a lack of progressive motility of spermatozoa from Mll5 deficient animals. None of these defects could be rescued by in vitro fertilization. Using microarray analysis we show that transcripts implicated in spermatogenesis are dysregulated. Our data demonstrate a clear role of Mll5 in mammalian spermatogenesis at the level of terminal differentiation providing further support for its classification in the SET3 branch of proteins. Moreover, this study identifies Tlk2, Utx, Gpr64, Sult4a1, Rap2ip, Vstm2 and HoxA10 as possible Mll5 targets that together may account for the observed spermatozoa maturation defects.


The outcomes of mandibular third molar removal and non-removal: a study of patients' preferences using a multi-attribute method.
The aim was to study patients' preferences about outcomes of mandibular third molar removal and non-removal using multi-attribute utility (MAU) methodology. The study comprised three stages. Stage 1: Elicitation of domains, i.e. main areas of patients' lives which could be affected by third molar removal and non-removal. Stage 2a: Interdomain weighting was obtained by relative weighting of the domains elicited in Stage 1. Stage 2b: Intradomain weighting obtained by patients' designation of values for different health states of each domain. Stage 3: Rating of outcomes. The patients were asked to imagine experiencing a variety of outcomes of mandibular third molar removal and non-removal, described in 19 short vignettes. The numbers of patients interviewed for the three stages were 30, 78, and 55, respectively. Five domains were identified. The mean relative weightings were approximately equal for the domains "Home and social life" and "General health and well-being", followed in order of importance by 'Job and studies" and "Health and comfort of mouth, teeth and gums". "Your appearance" received the lowest mean relative weighting. The vignette, which described the presence of a fluid-filled sac and suggested that this tooth must be removed, received the highest mean preference (least effect on patients' lives). The lowest mean preference (most effect on patients' lives) was generated by the vignette, which stated that the jaw was broken and that the teeth must be wired together for 6 weeks. We conclude that, from the patient's perspective, outcomes of non-removal were preferable to outcomes of mandibular third molar removal.


Pediatrics in an eighteenth century remedy book.
A manuscript home remedy book of several hundred pages in German, with entries from 1631 to 1861, was found in the Washington University School of Medicine Library some years ago. Two sections on diseases of children, from the early seventeenth to the mid-eighteenth centuries, have been translated and compared with both printed and manuscript pediatric works. People cited and geographicl places mentioned are discussed. The final conclusion reached is that the text is closer in tone and philosophy to the stream of manuscripts going back to classical times than it is to the printed book of its time. An appendix lists treatment of the wet-nurse in place of the child, and treatments for debility or wasting and pain in the abdomen, and traces these through classical and later manuscript sources.


The differentiation of HL-60 cells in the synthetic medium induced by GM3 ganglioside.
GM3 ganglioside, added exogenously to a promyelocytic leukemia cell line (HL-60 cells) in serum-free synthetic medium, induced differentiation into macrophage-like cells. Macrophagic morphology and function of differentiation-induced cells were determined by cell growth behavior, May-Grünwald-Giemsa staining, activities of nonspecific esterase, phagocytosis and nitroblue tetrazolium (NBT) reduction. GM3 ganglioside may play a role in triggering differentiation of HL-60 cells into macrophage-like cells.


New insights into the biogeochemistry of extremely acidic environments revealed by a combined cultivation-based and culture-independent study of two stratified pit lakes.
The indigenous microbial communities of two extremely acidic, metal-rich stratified pit lakes, located in the Iberian Pyrite Belt (Spain), were identified, and their roles in mediating transformations of carbon, iron, and sulfur were confirmed. A combined cultivation-based and culture-independent approach was used to elucidate microbial communities at different depths and to examine the physiologies of isolates, which included representatives of at least one novel genus and several species of acidophilic Bacteria. Phosphate availability correlated with redox transformations of iron, and this (rather than solar radiation) dictated where primary production was concentrated. Carbon fixed and released as organic compounds by acidophilic phototrophs acted as electron donors for acidophilic heterotrophic prokaryotes, many of which catalyzed the dissimilatory reduction in ferric iron; the ferrous iron generated was re-oxidized by chemolithotrophic acidophiles. Bacteria that catalyze redox transformations of sulfur were also identified, although these Bacteria appeared to be less abundant than the iron oxidizers/reducers. Primary production and microbial numbers were greatest, and biogeochemical transformation of carbon, iron, and sulfur, most intense, within a zone of c. 8-10 m depth, close to the chemocline, in both pit lakes. Archaea detected in sediments included two Thaumarchaeota clones, indicating that members of this recently described phylum can inhabit extremely acidic environments.


Neural detection of complex sound sequences in the absence of consciousness.
The neural response to a violation of sequences of identical sounds is a typical example of the brain's sensitivity to auditory regularities. Previous literature interprets this effect as a pre-attentive and unconscious processing of sensory stimuli. By contrast, a violation to auditory global regularities, i.e. based on repeating groups of sounds, is typically detectable when subjects can consciously perceive them. Here, we challenge the notion that global detection implies consciousness by testing the neural response to global violations in a group of 24 patients with post-anoxic coma (three females, age range 45-87 years), treated with mild therapeutic hypothermia and sedation. By applying a decoding analysis to electroencephalographic responses to standard versus deviant sound sequences, we found above-chance decoding performance in 10 of 24 patients (Wilcoxon signed-rank test, P < 0.001), despite five of them being mildly hypothermic, sedated and unarousable. Furthermore, consistently with previous findings based on the mismatch negativity the progression of this decoding performance was informative of patients' chances of awakening (78% predictive of awakening). Our results show for the first time that detection of global regularities at neural level exists despite a deeply unconscious state.


Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture.
Over the recent years, various materials have been introduced as potential 3D cell culture scaffolds. These include protein extracts, peptide amphiphiles, and synthetic polymers. Hydrogel scaffolds without human or animal borne components or added bioactive components are preferred from the immunological point of view. Here we demonstrate that native nanofibrillar cellulose (NFC) hydrogels derived from the abundant plant sources provide the desired functionalities. We show 1) rheological properties that allow formation of a 3D scaffold in-situ after facile injection, 2) cellular biocompatibility without added growth factors, 3) cellular polarization, and 4) differentiation of human hepatic cell lines HepaRG and HepG2. At high shear stress, the aqueous NFC has small viscosity that supports injectability, whereas at low shear stress conditions the material is converted to an elastic gel. Due to the inherent biocompatibility without any additives, we conclude that NFC generates a feasible and sustained microenvironment for 3D cell culture for potential applications, such as drug and chemical testing, tissue engineering, and cell therapy.


Formononetin protects neurons against hypoxia-induced cytotoxicity through upregulation of ADAM10 and sAβPPα.
Formononetin, an active constituent of the Chinese herb Astragali Radix, has been reported to have beneficial effects for Alzheimer's disease (AD). Yet the mechanism of this effect remains to be elucidated. The present study shows that formononetin increases soluble-AβPPα (sAβPPα) secretion and thus protects human-AβPP Swedish mutation cell (N2a-AβPP cell) from hypoxia-induced apoptosis. Using hypoxic N2a-AβPP cell as an in vitro model of AD-like pathology, we confirmed that regular treatment with formononetin could have neuroprotective effects, followed respectively by reduced caspase 3 activity and increased cell viability. Strikingly, our data revealed that the caspase 3-blocking effect of formononetin was largely mediated by stimulation of α-secretase cleavage of AβPP, and increasing the secretion of its soluble form, sAβPPα. Moreover, the protective effect of formononetin was totally inhibited by TAPI-2, an α-secretase complex inhibitor, suggesting the role of the sAβPPα pathway in the neuroprotective response to formononetin. We also found that the stimulative effect of formononetin on α-secretase activity was mainly conducted by upregulating ADAM10 expression at the transcriptional level. Altogether, our study provides novel insights into how formononetin mediates stimulation of the ADAM10-sAβPPα pathway and exerts a neuronal protective effect.


Investigating the Effectiveness and Acceptability of Oral Health and Related Health Behaviour Interventions in Adults with Severe and Multiple Disadvantage: Protocol for a Mixed-Methods Systematic Review.
Increasing numbers of people in England experience homelessness, substance use, and repeated offending (known as 'severe and multiple disadvantage'; SMD). Populations experiencing SMD often have extremely poor oral health, which is closely inter-linked with high levels of substance use, smoking, and poor diet. This study aims to undertake an evidence synthesis to identify the effectiveness, resource requirements, and factors influencing the implementation and acceptability of oral health and related health behaviour interventions in adults experiencing SMD. Two systematic reviews will be conducted using mixed-methods. Review 1 will investigate the effectiveness and resource implications of oral health and related health behaviours (substance use, smoking, diet) interventions; Review 2 will investigate factors influencing the implementation of such interventions. The population includes adults (≥18 years) experiencing SMD. Standard review methods in terms of searches, screening, data extraction, and quality appraisal will be conducted. Narrative syntheses will be conducted. If feasible, a meta-analysis will be conducted for Review 1 and a thematic synthesis for Review 2. Evidence from the two reviews will then be synthesised together. Input from people with experience of SMD will be sought throughout to inform the reviews. An initial logic model will be iteratively refined during the review.


Predicting history.
Can events be accurately described as historic at the time they are happening? Claims of this sort are in effect predictions about the evaluations of future historians; that is, that they will regard the events in question as significant. Here we provide empirical evidence in support of earlier philosophical arguments1 that such claims are likely to be spurious and that, conversely, many events that will one day be viewed as historic attract little attention at the time. We introduce a conceptual and methodological framework for applying machine learning prediction models to large corpora of digitized historical archives. We find that although such models can correctly identify some historically important documents, they tend to overpredict historical significance while also failing to identify many documents that will later be deemed important, where both types of error increase monotonically with the number of documents under consideration. On balance, we conclude that historical significance is extremely difficult to predict, consistent with other recent work on intrinsic limits to predictability in complex social systems2,3. However, the results also indicate the feasibility of developing 'artificial archivists' to identify potentially historic documents in very large digital corpora.


Experimental one-way quantum computing.
Standard quantum computation is based on sequences of unitary quantum logic gates that process qubits. The one-way quantum computer proposed by Raussendorf and Briegel is entirely different. It has changed our understanding of the requirements for quantum computation and more generally how we think about quantum physics. This new model requires qubits to be initialized in a highly entangled cluster state. From this point, the quantum computation proceeds by a sequence of single-qubit measurements with classical feedforward of their outcomes. Because of the essential role of measurement, a one-way quantum computer is irreversible. In the one-way quantum computer, the order and choices of measurements determine the algorithm computed. We have experimentally realized four-qubit cluster states encoded into the polarization state of four photons. We characterize the quantum state fully by implementing experimental four-qubit quantum state tomography. Using this cluster state, we demonstrate the feasibility of one-way quantum computing through a universal set of one- and two-qubit operations. Finally, our implementation of Grover's search algorithm demonstrates that one-way quantum computation is ideally suited for such tasks.


Predictive classified vector quantization.
A vector quantization scheme based on the classified vector quantization (CVQ) concept, called predictive classified vector quantization (PCVQ), is presented. Unlike CVQ where the classification information has to be transmitted, PCVQ predicts it, thus saving valuable bit rate. Two classifiers, one operating in the Hadamard domain and the other in the spatial domain, were designed and tested. The classification information was predicted in the spatial domain. The PCVQ schemes achieved bit rate reductions over the CVQ ranging from 20 to 32% for two commonly used color test images while maintaining the same acceptable image quality. Bit rates of 0.70-0.93 bits per pixel (bpp) were obtained depending on the image and PCVQ scheme used.


A Survey of County Health Departments of Kansas Regarding COVID-19.
SARS-CoV-2 virus (severe acute respiratory syndrome coronavirus 2) causing COVID-19 (Coronavirus Disease 2019) initially was identified in China in December 2019. It has resulted in a pandemic with increasing spread of the virus in the U.S. The county health departments around U.S. are spearheading the response to contain the spread of this virus. This project was a survey of county health departments in the state of Kansas with data collection period from April 15 to April 24, 2020. This study evaluated the staffing, resources, and funding of these health departments and how it was affecting the efforts to contain COVID-19. Descriptive statistics were used to summarize the responses. A total of 75% of the county health departments in Kansas responded to the survey. In 89% of locations, the staffing had not increased. Most health departments had an average of five people and the four largest ones had 30 to 98 staff working on COVID-19. Most locations used the Kansas Department of Health and Environment criteria for testing and used a combination of state or private laboratories. The results of the tests were available three days or longer in 62% and after five days in 14% of sites. All locations were active in contact tracing, but most had one to three people for this purpose and in 90% the contact tracing interview was via phone calls. There was no change in funding in 21% and decreased funding in 8.5% of health departments. Most locations had an average of five nasopharyngeal swabs on the day of the survey. The most common needs expressed were help to increase testing capability, more public education, more personal protective equipment, increased personnel, and assistance with contract tracing. There is an urgent need in Kansas to increase support to county health departments for testing capability, personal protective equipment, increased number of staff, increased help with contact tracing, and especially increase support for public education.


Diet is associated with ankle-brachial index, inflammation, and ambulation in patients with intermittent claudication.
The aims of this investigation were to determine whether the daily dietary intake of nutrients by patients with peripheral artery disease (PAD) and intermittent claudication (IC) met recommended levels for adults older than 50 years and to determine whether meeting recommended levels of nutrients was associated with ankle-brachial index (ABI), inflammation, and ambulation of patients with PAD and IC. A total of 48 patients were assessed on their dietary intake of 20 nutrients during a 3-day period. Patients were further characterized on demographic variables, comorbid conditions, cardiovascular risk factors, ABI, 6-minute walk distance (6MWD), and high-sensitivity C-reactive protein (hsCRP) concentration. Few patients met the daily recommended intakes for calcium (4%), fiber (6%), vitamin E (6%), trans fatty acids (13%), vitamin A (15%), total sugars (19%), potassium (23%), sodium (29%), saturated fat (29%), and vitamin C (31%), and none of the patients met the daily recommended intake of vitamin D (0%). Overall, patients met few of the 20 dietary recommendations as the median score was seven recommendations. Only 17 of 48 patients met more than seven of the recommendations. For the ABI regression model adjusted for age, sex, race, smoking, hypertension, dyslipidemia, body mass index, and percentage body fat, the only significant predictor was total sugars (P < .001); patients who did not meet the recommendation had lower ABI values. For the hsCRP-adjusted regression model, the strongest significant predictor was omega-3 polyunsaturated fatty acids (P = .001), indicating that those who did not meet the recommendation had higher hsCRP values. Finally, for the 6MWD-adjusted regression model, folate (P = .011) and dietary score index (P = .014) were significant predictors; those who did not meet the recommendation for folate and those who met 5 or fewer of the 20 recommendations had shorter 6MWD. Patients with PAD and IC consume a low-nutrient-dense diet that is deficient in many vitamins, calcium, fruits, and vegetables and contains too much added sugar, saturated and trans fats, and processed foods. In addition, more severe PAD, greater inflammation, and ambulatory dysfunction are independently associated with aspects of a low-nutrient-dense diet, such as too much intake of added sugars, low intake of omega-3 polyunsaturated fatty acids and folate, and meeting the recommended intakes of only five or fewer nutrients.


Indole acetic acid is responsible for protection against oxidative stress caused by drought in soybean plants: the role of heme oxygenase induction.
Objectives This study was focused on the role of indole acetic acid (IAA) in the defense against oxidative stress damage caused by drought in soybean plants and to elucidate whether heme oxygenase-1 (HO-1) and nitric oxide (NO) are involved in this mechanism. IAA is an auxin that participates in many plant processes including oxidative stress defense, but to the best of our knowledge no information is yet available about its possible action in drought stress. Methods To this end, soybean plants were treated with 8% polyethylene glycol (PEG) or 100 µM IAA. To evaluate the behavior of IAA, plants were pretreated with this compound previous to PEG addition. Lipid peroxidation levels (thiobarbituric acid reactive substances (TBARS)), glutathione (GSH) and ascorbate (AS) contents, catalase (CAT), superoxide dismutase (SOD), and guaiacol peroxidase (POD) activities were determined to evaluate oxidative damage. Results Drought treatment (8% PEG) caused a significant increase in TBARS levels as well as a marked decrease in the non-enzymatic (GSH and AS) and enzymatic (CAT, SOD, and POD) antioxidant defense systems. Pre-treatment with IAA prevented the alterations of stress parameters caused by drought, while treatment with IAA alone did not produce changes in TBARS levels, or GSH and AS contents. Moreover, the activities of the classical enzymes involved in the enzymatic defense system (SOD, CAT, and POD) remained similar to control values. Furthermore, this hormone could enhance HO-1 activity (75% with respect to controls), and this increase was positively correlated with protein content as well as gene expression. The direct participation of HO-1 as an antioxidant enzyme was established by performing experiments in the presence of Zn-protoporphyrin IX, a well-known irreversible inhibitor of this enzyme. It was also demonstrated that HO-1 is modulated by NO, as shown by experiments performed in the presence of an NO donor (sodium nitroprusside), an NO scavenger (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide), or an NO synthesis inhibitor (N-nitro-l-arginine methyl ester, NAME). Discussion It is concluded that IAA is responsible, at least in part, for the protection against oxidative stress caused by drought in soybean plants through the modulation of NO levels which, in turn, enhances HO-1 synthesis and activity.


Characteristics and outcomes among older HIV-positive adults enrolled in HIV programs in four sub-Saharan African countries.
Limited information exists on adults ≥50 years receiving HIV care in sub-Saharan Africa. Using routinely-collected longitudinal patient-level data among 391,111 adults ≥15 years enrolling in HIV care from January 2005-December 2010 and 184,689 initiating ART, we compared characteristics and outcomes between older (≥50 years) and younger adults at 199 clinics in Kenya, Mozambique, Rwanda, and Tanzania. We calculated proportions over time of newly enrolled and active adults receiving HIV care and initiating ART who were ≥50 years; cumulative incidence of loss to follow-up (LTF) and recorded death one year after enrollment and ART initiation, and CD4+ response following ART initiation. From 2005-2010, the percentage of adults ≥50 years newly enrolled in HIV care remained stable at 10%, while the percentage of adults ≥50 years newly initiating ART (10% [2005]-12% [2010]), active in follow-up (10% [2005]-14% (2010]), and active on ART (10% [2005]-16% [2010]) significantly increased. One year after enrollment, older patients had significantly lower incidence of LTF (33.1% vs. 32.6%[40-49 years], 40.5%[25-39 years], and 56.3%[15-24 years]; p-value<0.0001), but significantly higher incidence of recorded death (6.0% vs. 5.0% [40-49 years], 4.1% [25-39 years], and 2.8% [15-24 years]; p-valve<0.0001). LTF was lower after vs. before ART initiation for all ages, with older adults experiencing less LTF than younger adults. Among 85,763 ART patients with baseline and follow-up CD4+ counts, adjusted average 12-month CD4+ response for older adults was 20.6 cells/mm3 lower than for adults 25-39 years of age (95% CI: 17.1-24.1). The proportion of patients who are ≥50 years has increased over time and been driven by aging of the existing patient population. Older patients experienced less LTF, higher recorded mortality and less robust CD4+ response after ART initiation. Increased programmatic attention on older adults receiving HIV care in sub-Saharan Africa is warranted.


Approaches for the isolation and identification of hydrophilic, light-sensitive, volatile and minor natural products.
Covering: up to 2019The discovery of new bioactive natural products gained momentum during the last few decades, resulting from instrumentation advances, from the expansion of genome mining and regulation, as well as by exploration of untapped biological sources. However, water-soluble, volatile, minor and photosensitive natural products are yet poorly known. This review discusses the literature reporting the isolation strategies for some of these metabolites. Analysis of minor metabolites at sub-milligram level are also presented, since analytical instrumentation enabling structure assignment in minute quantities is now routine. Major trends related to natural products discovery are discussed, under the light of further developments in biodiscovery.


Indicators of Absolute and Relative Changes in Skeletal Muscle Mass during Adulthood and Ageing.
This study aimed to explore the set of variables related to skeletal muscle mass (SMM) in both sexes, and to create age- and sex-related models of changes in SMM, using the most representative indicator of muscular status. Body composition was assessed in 8733 subjects (♀ = 3370 and ♂ = 5363), allocated into subsamples according to age: 18-29.9, 30-39.9, 40-49.9, 50-59.9, 60-69.9, and 70.0-79.9 years. Nine variables were used: protein mass, protein percent, protein mass index, SMM, percent of SMM, SMM index, fat-free mass, fat-free mass index, and protein/fat index. Univariate and multivariate analysis of variance (ANOVA and MANOVA) were used to determine between- and within-sex difference in all variables by age. Correlation analysis established the relationship between age and muscularity variables. Principal Component Analysis extracted the variables that loaded highest in explaining muscularity, while regression analysis determined the linearity of association between the age and indicators of muscular status. Variables SMMI and PSMM were extracted as the most sensitive to age, with SMMI being gender-independent while showing the parabolic and sinusoidal form of change as function of ageing in males and females, respectively; and PSMM being sex-dependent while showing a linear trend of decrease in both sexes.


Effects of dietary fiber from wheat, corn, and soy hull bran on excretion of fecal bile acids in humans.
Effects of dietary fiber on bile acid excretion and fecal bile acid concentration have been studied for seven subjects fed 26 g of either soft white wheat bran, corn bran, soybean hulls, or hard red spring wheat bran. Results indicate that even in a controlled study using a metabolic word, individual subject variation has a major impact on fecal bile acid excretion. This observation has not been fully appreciated in previous human studies. No significant change in the composition of fecal bile acids could be associated with the decrease in serum lipid levels previously reported. A method for the isolation and quantitation of fecal bile acids is described which does not require purification by thin-layer chromatography. A preliminary study of lyophilized fecal samples stored at -10 to -30 degrees C showed very little or no change in bile acid content. Samples stored at room temperatures for 11 months showed a substantial reduction in bile acid content.


Breath analysis combined with cardiopulmonary exercise testing and echocardiography for monitoring heart failure patients: the AEOLUS protocol.
This paper describes the AEOLUS pilot study which combines breath analysis with cardiopulmonary exercise testing (CPET) and an echocardiographic examination for monitoring heart failure (HF) patients. Ten consecutive patients with a prior clinical diagnosis of HF with reduced left ventricular ejection fraction were prospectively enrolled together with 15 control patients with cardiovascular risk factors, including hypertension, type II diabetes or chronic ischemic heart disease. Breath samples were collected at rest and during CPET coupled with exercise stress echocardiography (CPET-ESE) protocol by means of needle trap micro-extraction and were analyzed through gas-chromatography coupled with mass spectrometry. The protocol also involved using of a selected ion flow tube mass spectrometer for a breath-by-breath isoprene and acetone analysis during exercise. At rest, HF patients showed increased breath levels of acetone and pentane, which are related to altered oxidation of fatty acids and oxidative stress, respectively. A significant positive correlation was observed between acetone and the gold standard biomarker NT-proBNP in plasma (r= 0.646,p< 0.001), both measured at rest. During exercise, some exhaled volatiles (e.g., isoprene) mirrored ventilatory and/or hemodynamic adaptation, whereas others (e.g., sulfide compounds and 3-hydroxy-2-butanone) depended on their origin. At peak effort, acetone levels in HF patients differed significantly from those of the control group, suggesting an altered myocardial and systemic metabolic adaptation to exercise for HF patients. These preliminary data suggest that concomitant acquisition of CPET-ESE and breath analysis is feasible and might provide additional clinical information on the metabolic maladaptation of HF patients to exercise. Such information may refine the identification of patients at higher risk of disease worsening.


Identification of the beta-binding domain of the alpha subunit of Escherichia coli polymerase III holoenzyme.
Rapid and processive DNA synthesis by Escherichia coli DNA polymerase III holoenzyme is achieved by the direct interaction between the alpha subunit of DNA polymerase III core and the beta sliding clamp (LaDuca, R. J., Crute, J. J., McHenry, C. S., and Bambara, R. A. (1986) J. Biol. Chem. 261, 7550-7557; Stukenberg, T. P., Studwell-Vaughan, P. S., and O'Donnell, M. (1991) J. Biol. Chem. 266, 11328-11334). In this study, we localized the beta-binding domain of alpha to a carboxyl-terminal region by quantifying the interaction of beta with a series of alpha deletion proteins. Purification and binding analysis was facilitated by insertion of hexahistidine and short biotinylation sequences on the deletion terminus of alpha. Interaction of beta with alpha deletion proteins was studied by gel filtration and surface plasmon resonance. alpha lacking 169 COOH-terminal residues still possessed beta-binding activity; whereas deletion of 342 amino acids from the COOH terminus abolished beta binding. Deletion of 542 amino acids from the NH2 terminus of the 1160 residue alpha subunit resulted in a protein that bound beta 10-20-fold more strongly than native alpha. Hence, portions of alpha between residues 542 and 991 are involved in beta binding. DNA binding to alpha apparently triggers an increased affinity for beta (Naktinis, V., Turner, J., and O'Donnell, M. (1996) Cell 84, 137-145). Our findings extend this observation by implicating the amino-terminal polymerase domain in inducing a low affinity taut conformation in the carboxyl-terminal beta-binding domain. Deletion of the polymerase domain (or, presumably, its occupancy by DNA) relaxes the COOH-terminal domain, permitting it to assume a conformation with high affinity for beta.


BCG vaccination impacts the epigenetic landscape of progenitor cells in human bone marrow.
While the Bacille-Calmette-Guérin (BCG) vaccine is used to prevent tuberculosis, it also offers protection against a diverse range of non-mycobacterial infections. However, the underlying protective mechanisms in humans are not yet fully understood. Here, we surveyed at single-cell resolution the gene expression and chromatin landscape of human bone marrow, aspirated before and 90 days after BCG vaccination or placebo administration. We show that BCG vaccination significantly alters both the gene expression and epigenetic profiles of human hematopoietic stem and progenitor cells (HSPCs). Changes in gene expression occur primarily on the most uncommitted stem cells and are reflective of a persistent myeloid bias. In contrast, BCG-induced changes in chromatin accessibility are most prevalent within differentiated progenitor cells at sites influenced by Kruppel-like factor (KLF)/SP and EGR transcription factors (TFs). These TFs are also activated in the most uncommitted stem cells, indicating that activated TFs, which drive persistent changes in HSC gene expression, likely also drive chromatin dynamics appearing within downstream progenitor cells. This perspective contests the prevailing notion that epigenetic modifications linked to innate immune memory transfer directly from stem cells to their differentiated derivatives. Finally, we show that alterations in gene expression and chromatin accessibility in HSPCs due to BCG vaccination were highly correlated (r>0.8) with the IL-1β secretion capacity of paired PBMCs upon secondary immune challenge. Overall, our findings shed light on BCG vaccination's profound and lasting effects on HSPCs and its influence on innate immune responses.


[Evaluation value of human antibacterial peptide LL-37 on the prognosis of elderly patients with sepsis].
To evaluate the prognostic value of human antibacterial peptide LL-37 in elderly patients with sepsis. Elderly sepsis patients over 65-year-old satisfied the diagnostic criteria for sepsis and septic shock admitted to intensive care unit of East Hospital of Tongji University from January 2016 to December 2017 were enrolled (elderly sepsis group). Aged community-acquired pneumonia (CAP) patients hospitalized during the same period were enrolled as a control group for pneumonia, and the aged health check-ups served as a healthy control group during the same period. The peripheral blood LL-37 levels of all patients on the 1st, 3rd, 7th day of admission and the results on the day of physical examination in the healthy control group and on the day of admission in aged CAP group were recorded. C-reactive protein (CRP), arterial blood lactate (Lac), procalcitonin (PCT) were monitored, and acute physiology and chronic health evaluation II (APACHE II) and sequential organ failure assessment (SOFA) scores were calculated based on the worst values within 24 hours. The correlation between LL-37 and various indicators was analyzed by Spearman method. According to the 28-day clinical outcome, the elderly patients with sepsis were divided into survival group and non-survival group. The differences in all parameters between the two groups were compared. The statistically significant indicators were analyzed by receiver operating characteristic (ROC) curve, and the predictive value of each indicator for prognosis was evaluated. (1) A total of 113 elderly patients with sepsis were enrolled in the final analysis, including 67 patients in sepsis group and 46 patients in septic shock group. Thirty-two patients were enrolled as healthy controls and 31 elderly patients with CAP as elderly pneumonia group. The PCT, CRP, Lac, APACHE II and SOFA scores of the patients in the three groups were higher than those of the healthy control group, and they were gradually increased with the severity of infection. There was no significant difference in gender or age among the groups. Compared with the healthy control group, the other three groups had higher LL-37 level after admission, the LL-37 levels in the sepsis group and the septic shock group were decreased with the prolongation of the hospitalization time, and they were lower than the pneumonia group at 7 days after admission [LL-37 (μg/L): 1 403.9±501.9, 1 517.1±676.4 vs. 1 608.4±816.2, both P > 0.05]. It was shown by correlation analysis that the LL-37 level in peripheral blood of elderly patients with sepsis was significantly negatively correlated with APACHE II score (r = -0.329, P = 0.007) and SOFA score (r = -0.344, P = 0.005), but no significant correlation with Lac was found (r = -0.128, P = 0.311). (2) The 28-day survival analysis revealed that of the 113 elderly patients with sepsis, 54 (47.8%) survived at 28 days and 59 (52.2%) died. There was no significant difference in gender, age, PCT or CRP levels at 1 day after admission between the two groups. The 1-day Lac, APACHE II and SOFA scores of the patients in the non-survival group were significantly higher than those in the survival group, they were gradually increased with the prolongation of the hospitalization time, and they were significantly higher than those in the survival group at 7 days after admission [Lac (mmol/L): 2.4 (1.4, 4.4) vs. 1.0 (0.8, 1.7), APACHE II score: 21.77±5.85 vs. 13.74±4.99, SOFA score: 9.62±4.78 vs. 3.18±2.71, all P < 0.01]. With the prolongation of admission, there was no significant change in LL-37 level of peripheral blood in the survival group. The LL-37 level in the non-survival group showed a downward tendency, and it was significantly lower than that in the survival group at 7 days after admission (μg/L: 1 277.8±642.6 vs. 1 620.6±461.6, P < 0.05). It was shown by ROC curve analysis that the LL-37 in peripheral blood, Lac, APACHE II score and SOFA score at 7-day of admission of elderly patients with sepsis had predictive value for prognosis, and LL-37 had the best predicted effect for 28-day death, the area under the ROC curve (AUC) of LL-37 was 0.670, 95% confidence interval (95%CI) = 0.513-0.757, when the optimal cut-off value was 1 283.0 μg/L, the sensitivity was 75.7%, and the specificity was 61.5%. The expression of LL-37 increased in the early course of the disease in elderly patients with sepsis. However, as the disease progressed and worsened, the level of LL-37 had a decline tendency and was associated with death. The dynamic monitoring of LL-37 combined with APACHE II and SOFA scores had clinical guidance value in predicting the prognosis of sepsis in the elderly.


α-Kleisin subunit of cohesin preserves the genome integrity of embryonic stem cells.
Cohesin is a ring-shaped protein complex that comprises the SMC1, SMC3, and α-kleisin proteins, STAG1/2/3 subunits, and auxiliary factors. Cohesin participates in chromatin remodeling, chromosome segregation, DNA replication, and gene expression regulation during the cell cycle. Mitosis-specific α-kleisin factor RAD21 and meiosis-specific α-kleisin factor REC8 are expressed in embryonic stem cells (ESCs) to maintain pluripotency. Here, we demonstrated that RAD21 and REC8 were involved in maintaining genomic stability and modulating chromatin modification in murine ESCs. When the kleisin subunits were depleted, DNA repair genes were downregulated, thereby reducing cell viability and causing replication protein A (RPA) accumulation. This finding suggested that the repair of exposed single-stranded DNA was inefficient. Furthermore, the depletion of kleisin subunits induced DNA hypermethylation by upregulating DNA methylation proteins. Thus, we proposed that the cohesin complex plays two distinct roles in chromatin remodeling and genomic integrity to ensure the maintenance of pluripotency in ESCs. [BMB Reports 2023; 56(2): 108-113].


[Factors associated with use of oral health services: a population-based study in municipalities of the state of Maranhão, Brazil].
The scope of this study was to evaluate the use of oral health services (OHS) and related factors in municipalities with more than 100,000 inhabitants in Maranhão, Brazil. It was a population-based sample including 1214 children and 1059 adults. Prevalence ratios (PR) were estimated by hierarchical Poisson regression in accordance with Andersen's theoretical model. Over 91% of children and 71.9% of adults had not used the OHS in the six months prior to the interview. Of those who did, 48.5% were attended in the Unified Health System (SUS). Preventive were more frequent than curative procedures. In children and adults, predisposing, facilitating and need-based factors explained the use of OHS. Children aged >2 years (PR=5.29), with greater schooling of the head of the household (PR=2.37), e"6 prenatal visits (PR=1.69) and dental treatment needs (PR=9.54) were associated with greater use of the OHS. In adults, use was associated with greater schooling (PR=2.26), economic class A/B (PR=1.38), self-perceived health as good/very good (PR=1.72) and need for treatment (PR=18.25). The use of the OHS is neither universal nor equitable and there are deficiencies in comprehensive care, as few people use the more complex services. Fewer prenatal visits appear to be a predictor of non-use of the OHS by children.


Impact of a randomized, controlled trial of liberal vs conservative hospital discharge criteria on energy, protein, and fluid intake in patients who received marrow transplants.
To determine if adult patients who received marrow transplants had faster resumption of oral energy and nutrient intake and shorter duration of intravenous (i.v.) fluid requirement if discharged from the hospital earlier than is customary. Randomized, controlled trial of patients remaining hospitalized because of inadequate oral intake. Consenting patients were assigned randomly to remain hospitalized (hospital group) or be discharged to an ambulatory setting (ambulatory group). Seventy-eight patients of the Fred Hutchinson Cancer Research Center who were consuming less than 33% of estimated energy requirement and requiring up to 3,000 mL of fluids per day intravenously. Participants received nutrition counseling by a registered dietitian to promote resumption of oral intake. Daily oral intake records were analyzed to determine energy and nutrient content. Days after study enrollment to consume 33% of energy and protein requirements and total number of days of i.v. fluid support were analyzed by group until discharge from the center, approximately 100 days after transplantation. Demographic data were defined by group means. Differences between treatment procedures were determined by Cox regression analysis. No variables were confounding. The hospital group took fewer days than the ambulatory group to resume oral energy intake (4.5 vs 8.0, P = .004) and to discontinue i.v. fluids (30.5 vs 48.5, P = .019). There was no difference between groups in days of parenteral nutrition support (P = .817) or days to resume oral protein intake (P = .470). Oral and gastrointestinal complications delay resumption of oral energy and protein intakes after transplantation. Earlier hospital discharge can achieve cost savings but may delay resumption of oral energy intake. Because of continued high-risk nutrition status and potential for rapid change in medical status, nutrition assessment and counseling are necessary in both the hospital and ambulatory setting to promote resumption of oral intake and discontinuation of i.v. fluids.


Developing a novel international undergraduate surgical masterclass during a financial crisis: our 4-year experience.
Essential Skills in the Management of Surgical Cases (ESMSC) is a novel 3-day international undergraduate surgical masterclass. Its current curriculum (Cores integrated for Research-Ci4R) is built on a tetracore, multiclustered architecture combining high-fidelity and low-fidelity simulation-based learning (SBL), with applied and basic science case-based workshops, and non-technical skills modules. We aimed to report our experience in setting up ESMSC during the global financial crisis. We report the evolution of our curriculum's methodology and summarised the research outcomes related to the objective performance improvement of delegates, the educational environment of the course and the use of mixed-fidelity SBL. Feedback from the last three series of the course was prospectively collected and analysed using univariate statistics on IBM SPSS V.23. 311 medical students across the European Union (EU) were selected from a competitive pool of 1280 applicants during seven series of the course between 2014 and 2017. During this period, curriculum 14 s evolved to the final Ci4R version, which integrates a tetracore structure combining 32 stations of in vivo, ex vivo and dry lab SBL with small group teaching workshops. Ci4R was positively perceived across different educational background students (p>0.05 for any comparison). ESMSC is considered an innovative and effective multidisciplinary teaching model by delegates, where it improves delegates objective performance in basic surgical skills. Our experience demonstrates provision of high-quality and free surgical education during a financial crisis, which evolved through a dynamic feedback mechanism. The prospective recording and subsequent analysis of curriculum evolution provides a blueprint to direct development of effective surgical education courses that can be adapted to local needs.


Development and evaluation of a survey instrument to assess veterinary medical record suitability for multi-center research studies.
Here we describe the development and evaluation of a survey instrument to assess the research suitability of veterinary electronic medical records (EMRs) through the conduct of two studies as part of the Dog Aging Project (DAP). In study 1, four reviewers used the instrument to score a total of 218 records in an overlapping matrix of pairs to assess inter-rater agreement with respect to appropriate format (qualification), identification match (verification), and record quality. Based upon the moderate inter-rater agreement with respect to verification and the relatively large number of records that were incorrectly rejected the instrument was modified and more specific instructions were provided. In study 2, a modified instrument was again completed by four reviewers to score 100 different EMRs. The survey scores were compared to a gold standard of board-certified specialist review to determine receiver operating curve statistics. The refined survey had substantial inter-rater agreement across most qualification and verification questions. The cut-off value identified had a sensitivity of 95 and 96% (by reviewer 1 and reviewer 2, respectively) and a specificity of 82% and 91% (by reviewer 1 and reviewer 2, respectively) to predict gold standard acceptance or rejection of the record. Using just qualification and verification questions within the instrument (as opposed to full scoring) minimally impacted sensitivity and specificity and resulted in substantial time savings in the review process.


Long-term deterioration of quality of life in adult patients with celiac disease is associated with treatment noncompliance.
Deterioration of quality of life in the long term has been suggested for celiac disease patients on a gluten-free diet. To determine long-term quality of life of celiac disease patients and to assess the benefits of gluten-free diet compliance. We prospectively evaluated 53 newly diagnosed adult celiac disease patients. The Short Form 36 Health Survey, the Gastrointestinal Symptoms Rating Scale and the Beck Depression Inventory were employed at the time of diagnosis, 1 year, and beyond 4 years (median: 53 months) on treatment. At 1 year, a significant improvement from baseline in quality of life indicators was observed (p<0.001 to p<0.0001) with comparable scores to healthy subjects. At 4 years, the Short Form 36 Health Survey scores (p<0.002 to p<0.0002) and Beck Depression Inventory score (p<0.002) show significant deterioration compare with 1 year. Most scores remained significantly better than those at diagnosis (p<0.03 to p<0.0005). No changes were detected in the Gastrointestinal Symptoms Rating Scale scores. The long-term impairment of quality of life was attributable to the deterioration of most dimensions in patients who were not strictly compliant with the gluten-free diet (p<0.05 to p<0.001). Long-term deterioration of quality of life outcomes after the first year of gluten-free diet was associated with the lack of strict compliance with the diet.


Association study of 25 type 2 diabetes related Loci with measures of obesity in Indian sib pairs.
Obesity is an established risk factor for type 2 diabetes (T2D) and they are metabolically related through the mechanism of insulin resistance. In order to explore how common genetic variants associated with T2D correlate with body mass index (BMI), we examined the influence of 25 T2D associated loci on obesity risk. We used 5056 individuals (2528 sib-pairs) recruited in Indian Migration Study and conducted within sib-pair analysis for six obesity phenotypes. We found associations of variants in CXCR4 (rs932206) and HHEX (rs5015480) with higher body mass index (BMI) (β=0.13, p=0.001) and (β=0.09, p=0.002), respectively and weight (β=0.13, p=0.001) and (β=0.09, p=0.001), respectively. CXCR4 variant was also strongly associated with body fat (β=0.10, p=0.0004). In addition, we demonstrated associations of CXCR4 and HHEX with overweight/obesity (OR=1.6, p=0.003) and (OR=1.4, p=0.002), respectively, in 1333 sib-pairs (2666 individuals). We observed marginal evidence of associations between variants at six loci (TCF7L2, NGN3, FOXA2, LOC646279, FLJ39370 and THADA) and waist hip ratio (WHR), BMI and/or overweight which needs to be validated in larger set of samples. All the above findings were independent of daily energy consumption and physical activity level. The risk score estimates based on eight significant loci (including nominal associations) showed associations with WHR and body fat which were independent of BMI. In summary, we establish the role of T2D associated loci in influencing the measures of obesity in Indian population, suggesting common underlying pathophysiology across populations.