Novel Role for the AnxA1-Fpr2/ALX Signaling Axis as a Key Regulator of Platelet Function to Promote Resolution of Inflammation.
Ischemia reperfusion injury (I/RI) is a common complication of cardiovascular diseases. Resolution of detrimental I/RI-generated prothrombotic and proinflammatory responses is essential to restore homeostasis. Platelets play a crucial part in the integration of thrombosis and inflammation. Their role as participants in the resolution of thromboinflammation is underappreciated; therefore we used pharmacological and genetic approaches, coupled with murine and clinical samples, to uncover key concepts underlying this role. Middle cerebral artery occlusion with reperfusion was performed in wild-type or annexin A1 (AnxA1) knockout (AnxA1-/-) mice. Fluorescence intravital microscopy was used to visualize cellular trafficking and to monitor light/dye-induced thrombosis. The mice were treated with vehicle, AnxA1 (3.3 mg/kg), WRW4 (1.8 mg/kg), or all 3, and the effect of AnxA1 was determined in vivo and in vitro. Intravital microscopy revealed heightened platelet adherence and aggregate formation post I/RI, which were further exacerbated in AnxA1-/- mice. AnxA1 administration regulated platelet function directly (eg, via reducing thromboxane B2 and modulating phosphatidylserine expression) to promote cerebral protection post-I/RI and act as an effective preventative strategy for stroke by reducing platelet activation, aggregate formation, and cerebral thrombosis, a prerequisite for ischemic stroke. To translate these findings into a clinical setting, we show that AnxA1 plasma levels are reduced in human and murine stroke and that AnxA1 is able to act on human platelets, suppressing classic thrombin-induced inside-out signaling events (eg, Akt activation, intracellular calcium release, and Ras-associated protein 1 [Rap1] expression) to decrease αIIbβ3 activation without altering its surface expression. AnxA1 also selectively modifies cell surface determinants (eg, phosphatidylserine) to promote platelet phagocytosis by neutrophils, thereby driving active resolution. (n=5-13 mice/group or 7-10 humans/group.) Conclusions: AnxA1 affords protection by altering the platelet phenotype in cerebral I/RI from propathogenic to regulatory and reducing the propensity for platelets to aggregate and cause thrombosis by affecting integrin (αIIbβ3) activation, a previously unknown phenomenon. Thus, our data reveal a novel multifaceted role for AnxA1 to act both as a therapeutic and a prophylactic drug via its ability to promote endogenous proresolving, antithromboinflammatory circuits in cerebral I/RI. Collectively, these results further advance our knowledge and understanding in the field of platelet and resolution biology.


Less is more: Independent loss-of-function OCIMENE SYNTHASE alleles parallel pollination syndrome diversification in monkeyflowers (Mimulus).
Pollinator-mediated selection on flower phenotypes (e.g., shape, color, scent) is key to understanding the adaptive radiation of angiosperms, many of which have evolved specialized relationships with a particular guild of animal pollinators (e.g., birds, bats, moths, bees). E-β-Ocimene, a monoterpene produced by OCIMENE SYNTHASE (OS) in Mimulus lewisii, is a floral scent important in attracting the species' bumblebee pollinators. The taxa closely related to M. lewisii have evolved several different pollination syndromes, including hummingbird pollination and self pollination (autogamy). We are interested in how floral scent variation contributed to species diversification in this clade. We analyzed variation in E-β-ocimene emission within this Mimulus clade and explored its molecular basis through a combination of DNA sequencing, reverse transcriptase PCR, and enzyme functional analysis in vitro. We found that none of the taxa, other than M. lewisii, emitted E-β-ocimene from flowers. But the molecular basis underlying loss of E-β-ocimene emission is unique in each taxon, including deletion, missense, or frameshift mutations in the OS gene, and potential posttranscriptional downregulation. The molecular evidence suggests that parallel loss-of-function in OS is the best explanation for the observed pattern of E-β-ocimene emission, likely as the result of natural selection.


Assessing capacity to consent for research in cognitively impaired older patients.
The number of clinical trials including older patients, and particularly patients with cognitive impairment, is increasing. While statutory provisions exist to make sure that the capacity to consent is assessed systematically for each patient, many gray areas remain with regard to how this assessment is made or should be made in the routine practice of clinical research. The aim of this review was to draw up an inventory of assessment tools evaluating older patients' capacity to consent specifically applicable to clinical research, which could be used in routine practice. Two authors independently searched PubMed, Cochrane, and Google Scholar data-bases between November 2015 and January 2016. The search was actualized in April 2017. We used keywords (MeSH terms and text words) referring to informed consent, capacity to consent, consent for research, research ethics, cognitive impairment, vulnerable older patients, and assessment tools. Existing reviews were also considered. Among the numerous existing tools for assessing capacity to consent, 14 seemed potentially suited for clinical research and six were evaluated in older patients. The MacArthur Competence Assessment Tool for Clinical Research (MacCAT-CR) was the most frequently cited. The MacCAT-CR is currently the most used and the best validated questionnaire. However, it appears difficult to use and time-consuming. A more recent tool, the University of California Brief Assessment of Capacity to Consent (UBACC), seems interesting for routine practice because of its simplicity, relevance, and applicability in older patients.


Cytosolic Ca2+ transients during pulsed focused ultrasound generate reactive oxygen species and cause DNA damage in tumor cells.
Mechanical forces from non-ablative pulsed focused ultrasound (pFUS) generate pro-inflammatory tumor microenvironments (TME), marked by increased cytokines, chemokines, and trophic factors, as well as immune cell infiltration and reduced tumor growth. pFUS also causes DNA damage within tumors, which is a potent activator of immunity and could contribute to changes in the TME. This study investigated mechanisms behind the mechanotransductive effects of pFUS causing DNA damage in several tumor cell types. Methods: 4T1 (murine breast tumor), B16 (murine melanoma), C6 (rat glioma), or MDA-MB-231 (human breast tumor) cells were sonicated in vitro (1.1MHz; 6MPa PNP; 10ms pulses; 10% duty cycle; 300 pulses). DNA damage was detected by TUNEL, apoptosis was measured by immunocytochemistry for cleaved caspase-3. Calcium, superoxide, and H2O2 were detected by fluorescent indicators and modulated by BAPTA-AM, mtTEMPOL, or Trolox, respectively. Results: pFUS increased TUNEL reactivity (range = 1.6-2.7-fold) in all cell types except C6 and did not induce apoptosis in any cell line. All lines displayed cytosolic Ca2+ transients during sonication. pFUS increased superoxide (range = 1.6-2.0-fold) and H2O2 (range = 2.3-2.8-fold) in all cell types except C6. BAPTA-AM blocked increased TUNEL reactivity, superoxide and H2O2 formation, while Trolox also blocked increased TUNEL reactivity increased after pFUS. mtTEMPOL allowed H2O2 formation and did not block increased TUNEL reactivity after pFUS. Unsonicated C6 cells had higher baseline concentrations of cytosolic Ca2+, superoxide, and H2O2, which were not associated with greater baseline TUNEL reactivity than the other cell lines. Conclusions: Mechanotransduction of pFUS directly induces DNA damage in tumor cells by cytosolic Ca2+ transients causing formation of superoxide and subsequently, H2O2. These results further suggest potential clinical utility for pFUS. However, the lack of pFUS-induced DNA damage in C6 cells demonstrates a range of potential tumor responses that may arise from physiological differences such as Ca2+ or redox homeostasis.


Coronary plaque imaging by coronary computed tomography angiography.
Coronary computed tomography angiography (CTA) has become the useful noninvasive imaging modality alternative to the invasive coronary angiography for detecting coronary artery stenoses in patients with suspected coronary artery disease (CAD). With the development of technical aspects of coronary CTA, clinical practice and research are increasingly shifting toward defining the clinical implication of plaque morphology and patients outcomes by coronary CTA. In this review we discuss the coronary plaque morphology estimated by CTA beyond coronary angiography including the comparison to the currently available other imaging modalities used to examine morphological characteristics of the atherosclerotic plaque. Furthermore, this review underlies the value of a combined assessment of coronary anatomy and myocardial perfusion in patients with CAD, and adds to an increasing body of evidence suggesting an added diagnostic value when combining both modalities. We hope that an integrated, multi-modality imaging approach will become the gold standard for noninvasive evaluation of coronary plaque morphology and outcome data in clinical practice.


Genealogic origins of nurse-midwifery education programs in the United States.
This article presents a genealogy of all known basic and refresher nurse-midwifery education programs enabling every CNM and CM to track their individual lineage back to Hattie Hemschemeyer or Mary Breckinridge. Feeling connected to our founding foremothers increases our understanding of who we are and what our commitment is to the families we serve. Genealogy also gives us an opportunity to reflect on the early history and continuing historical trends of our education programs.


[Trisomy 4p as result of a maternal translocation t(4;8)(q11;p23)].
We report on a 21 month old boy with congenital anomalies and a trisomy of the short arm of chromosome 4 as result of a maternal translocation 4/8 with meiotic 3:1 segregation. Karyotype/phenotype correlation is compared with other reported cases and discussed in a context of nearly pure trisomy 4p.


Comparative performance of modern digital mammography systems in a large breast screening program.
To compare physical measures pertaining to image quality among digital mammography systems utilized in a large breast screening program. To examine qualitatively differences in these measures and differences in clinical cancer detection rates between CR and DR among sites within that program. As part of the routine quality assurance program for screening, field measurements are made of several variables considered to correlate with the diagnostic quality of medical images including: modulation transfer function, noise equivalent quanta, d' (an index of lesion detectability) and air kerma to allow estimation of mean glandular dose. In addition, images of the mammography accreditation phantom are evaluated. It was found that overall there were marked differences between the performance measures of DR and CR mammography systems. In particular, the modulation transfer functions obtained with the DR systems were found to be higher, even for larger detector element sizes. Similarly, the noise equivalent quanta, d', and the phantom scores were higher, while the failure rates associated with low signal-to-noise ratio and high dose were lower with DR. These results were consistent with previous findings in the authors' program that the breast cancer detection rates at sites employing CR technology were, on average, 30.6% lower than those that used DR mammography. While the clinical study was not large enough to allow a statistically powered system-by-system assessment of cancer detection accuracy, the physical measures expressing spatial resolution, and signal-to-noise ratio are consistent with the published finding that sites employing CR systems had lower cancer detection rates than those using DR systems for screening mammography.


Risk factors for diabetic choroidopathy in patients with diabetic retinopathy.
To identify risk factors for diabetic choroidopathy in patients with type 2 diabetes. Forty-five consecutive patients with diabetic retinopathy underwent simultaneous indocyanine green angiography (ICG) and fluorescein angiography using a double detector and confocal scanning laser ophthalmoscopy in both eyes. Choroidal vascular abnormalities were evaluated by comparing the angiographic findings derived from the two methods. We analyzed the association between the presence of abnormal choroidal lesions evident on ICG angiography and several risk factors. Choroidal abnormalities evident on ICG angiography but not on fluorescein angiography included hypofluorescent spots in 72 eyes (80%) of 40 patients (89%), small hyperfluorescent spots in 61 eyes (68%) of 35 patients (78%), and large hyperfluorescent spots in 32 eyes (36%) of 21 patients (47%). The severity of diabetic retinopathy was significantly associated with the presence of hypofluorescent spots ( P=0.002, Cochran-Armitage test) in both eyes and with the small hyperfluorescent spots in the right eyes ( P=0.047, Cochran-Armitage test). Glycosylated hemoglobin levels were significantly associated with the large hyperfluorescent spots in the right eyes ( P=0.003, Fisher's exact probability test), and the treatment regimen was significantly associated with the small hyperfluorescent spots in the left eyes ( P=0.048, chi-square test). These data suggest that risk factors influencing the prevalence of diabetic choroidopathy in patients with type 2 diabetes mellitus may include severity of diabetic retinopathy, degree of diabetic control and treatment regimen.


A study of evaluating specific tissue oxygen saturation values of gastrointestinal tumors by removing adherent substances in oxygen saturation imaging.
Oxygen saturation (OS) imaging is a new method of endoscopic imaging that has clinical applications in oncology which can directly measure tissue oxygen saturation (Sto2) of the surface of gastrointestinal tract without any additional drugs or devices. This imaging technology is expected to contribute to research into cancer biology which leads to clinical benefit such as prediction to efficacy of chemotherapy or radiotherapy. However, adherent substances on tumors such as blood and white coating, pose a challenge for accurate measurements of the StO2 values in tumors. The aim of this study was to develop algorithms for discriminating between the tumors and their adherent substances, and to investigate whether it is possible to evaluate the tumor specific StO2 values excluding adherent substances during OS imaging. We plotted areas of tumors and their adherent substances using white-light images of 50 upper digestive tumors: blood (68 plots); reddish tumor (83 plots); white coating (89 plots); and whitish tumor (79 plots). Scatter diagrams and discriminating algorithms using spectrum signal intensity values were constructed and verified using validation datasets. StO2 values were compared between the tumors and tumor adherent substances using OS images of gastrointestinal tumors. The discriminating algorithms and their accuracy rates (AR) were as follows: blood vs. reddish tumor: Y> - 4.90X+7.13 (AR: 95.9%) and white coating vs. whitish tumor: Y< -0.52X+0.17 (AR: 96.0%). The StO2 values (median, [range]) were as follows: blood, 79.3% [37.8%-100.0%]; reddish tumor, 74.5% [62.0%-86.9%]; white coating, 73.8% [42.1%-100.0%]; and whitish tumor, 65.7% [53.0%-76.3%]. OS imaging is strongly influenced by adherent substances for evaluating the specific StO2 value of tumors; therefore, it is important to eliminate the information of adherent substances for clinical application of OS imaging.


Mesoporous structures confined in anodic alumina membranes.
The synthesis of nanoporous membranes based on different concepts and materials is a field of active research. This review focuses on the synthesis strategies, mesophase evolution mechanisms and potential applications of mesoporous materials confined within anodic alumina membranes (AAM). Following a rapid evolution of synthetic techniques, a significant number of different mesoporous materials (e.g., silica, titania, and carbon) with highly regular structures can now be prepared within these membranes. In recent years, efforts have also been made to understand the formation mechanisms of these hierarchical mesophases. The resulting organized nanoporous membranes open up a wide range of potential applications in fields such as templating oriented nanowires and controlled separation and release of molecules. For example, while various synthesis strategies can be used for the preparation of membrane-embedded nanowires, the latter can also be obtained as isolated objects after dissolution of the alumina host matrix. The review also discusses issues such as control of structural defects or integrity of interfaces that should be addressed in future research in order to fully exploit the potential of these hierarchical mesoporous channel structures.


Drosophila nipped-B protein supports sister chromatid cohesion and opposes the stromalin/Scc3 cohesion factor to facilitate long-range activation of the cut gene.
The Drosophila melanogaster Nipped-B protein facilitates transcriptional activation of the cut and Ultrabithorax genes by remote enhancers. Sequence homologues of Nipped-B, Scc2 of Saccharomyces cerevisiae, and Mis4 of Schizosaccharomyces pombe are required for sister chromatid cohesion during mitosis. The evolutionarily conserved Cohesin protein complex mediates sister chromatid cohesion, and Scc2 and Mis4 are needed for Cohesin to associate with chromosomes. Here, we show that Nipped-B is also required for sister chromatid cohesion but that, opposite to the effect of Nipped-B, the stromalin/Scc3 component of Cohesin inhibits long-range activation of cut. To explain these findings, we propose a model based on the chromatin domain boundary activities of Cohesin in which Nipped-B facilitates cut activation by alleviating Cohesin-mediated blocking of enhancer-promoter communication.


Safety and efficacy of 5-aminolevulinic acid phosphate/iron in mild-to-moderate coronavirus disease 2019: A randomized exploratory phase II trial.
5-aminolevulinic acid (5-ALA), a natural amino acid that is marketed alongside sodium ferrous citrate (SFC) as a functional food, blocks severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) proliferation in vitro and exerts anti-inflammatory effects. In this phase II open-label, prospective, parallel-group, randomized trial, we aimed to evaluate the safety and efficacy of 5-ALA in patients with mild-to-moderate coronavirus disease 2019. This trial was conducted in patients receiving 5-ALA/SFC (250/145 mg) orally thrice daily for 7 days, followed by 5-ALA/SFC (150/87 mg) orally thrice daily for 7 days. The primary endpoints were changes in SARS-CoV-2 viral load, clinical symptom scores, and 5-ALA/SFC safety (adverse events [AE] and changes in laboratory values and vital signs). A total of 50 patients were enrolled from 8 institutions in Japan. The change in SARS-CoV-2 viral load from baseline was not significantly different between the 5-ALA/SFC (n = 24) and control (n = 26) groups. The duration to improvement was shorter in the 5-ALA/SFC group than in the control group, although the difference was not significant. The 5-ALA/SFC group exhibited faster improvement rates in "taste abnormality," "cough," "lethargy," and "no appetite" than the control group. Eight AEs were observed in the 5-ALA/SFC group, with 22.7% of patients experiencing gastrointestinal symptoms (decreased appetite, constipation, and vomiting). AEs occurred with 750/435 mg/day in 25.0% of patients in the first phase and with 450/261 mg/day of 5-ALA/SFC in 6.3% of patients in the second phase. 5-ALA/SFC improved some symptoms but did not influence the SARS-CoV-2 viral load or clinical symptom scores over 14 days. The safety of 5-ALA/SFC in this study was acceptable. Further evaluation using a larger sample size or modified method is warranted.


Length of initial prescription at hospital discharge and long-term medication adherence for elderly patients with coronary artery disease: a population-level study.
Patient adherence to cardiac secondary prevention medications declines over time. We examined whether the length of the initial prescription at hospital discharge after coronary angiography would be associated with long-term adherence. We conducted a population-level cohort study to examine adherence to cardiac medications for 18 months after coronary angiography in elderly patients with coronary artery disease (CAD). We identified patients with clinical indications for angiotensin-converting enzyme inhibitors or angiotensin receptor blockers (ACE-I/ARB), beta-blockers (BB), and/or statins. In each medication class cohort, we defined high adherence as proportion of days covered (PDC) > 80%. The length of the initial prescription was defined as 0-30 days, 31-60 days, and more than 60 days. We controlled for patient sociodemographic factors, previous adherence, and comorbidities. The ACE-I/ARB cohort included 13,305 patients, the BB cohort included 5,792 patients, and the statin cohort included 16,134 patients. Using < 30 days as the reference, initial prescriptions covering at least 60 days were more likely to result in high long-term adherence for ACE-I/ARB (adjusted odds ratio [aOR], 4.1; 95% confidence interval [CI], 3.6-4.7); BB (aOR, 2.4; 95% CI, 1.9-3.1), and statins (aOR, 3.0; 95% CI, 2.6-3.4). More than 80% of patients had outpatient follow-up with a primary care provider within 30 days, and this did not vary based on length of initial prescription. Giving patients longer prescriptions for cardiac secondary prevention medications at hospital discharge seems to increase the likelihood of high long-term adherence in elderly patients.


Getting to the Heart of the Matter: Navigating Narrative Intersections in Ethics Consultation.
Ethics consultants can apply a narrative ethics approach to address ethical challenges that arise in critical situations. This approach recognizes how those involved in the narrative make sense of, keep faith with, and try on new identities and new understanding of their stories. This case study explores the ways in which the stories of patient, provider, and clinical ethics consultant intersect, and considers how the organic nature of the narrative ethics approach allows ethics consultants to navigate the stories of multiple stakeholders as they grapple with complex health care decisions. This essay also suggests that clinical ethics consultants applying the lens of narrative ethics have an obligation to approach consultations with courage, professional humility, intellectual curiosity, and an appreciation for the narratives of as many of the stakeholders as possible (including one's own).


Renal sodium excretion and the peritubular capillary physical factors in essential hypertension.
Peritubular capillary hydrostatic and oncotic forces and their relationship to the renal excretion of sodium (UNaV) were examined in 19 patients with moderate and uncomplicated essential hypertension (HT) and compared with data from 20 normotensive subjects (NT). Observations were made in hydropenia (C) and during sustained isotonic saline volume expansion (E; 3% increase in body weight). The intrarenal venous pressure (IRVP) was used as an index of peritubular capillary hydrostatic pressure, and the efferent arteriolar colloid osmotic pressure (COPeff) was estimated from the arterial COP and the filtration fraction. C values (mean +/- SEM) in HT (and NT) were: arterial pressure (MAP) 110 +/- 3 mm Hg (85 +/- 1, p less than 0.001); glomerular filtration rate (GFR) 122 +/- 4 ml/min/1.73 m2 (128 +/- 3, p greater than 0.05); renal blood flow (RBF) 1172 +/- 38 ml/min/1.73 m2 (1298 +/- 48, p less than 0.05); IRVP 25.0 +/- 1.0 mm Hg (24.8 +/- 0.8, p greater than 0.05); COPeff 33.0 +/- 0.7 mm Hg (31.9 +/- 0.6, p greater than 0.05); and UNaV 140 +/- 13 mumole/min (161 +/- 12, p greater than 0.05). During E, the increase of UNaV in HT was more than double that of NT (p less than 0.001) while IRVP did not change in either group (p greater than 0.05); and COPeff fell by 26% (p less than 0.001) in both groups. GFR and RBF increased by 18% (p less than 0.001) and 19% (p less than 0.001) respectively, in HT, but did not change in NT. MAP remained unchanged in both groups. The results indicate that the peritubular capillary physical factors are normal in established essential hypertension, and that these forces are not involved in the exaggerated natriuretic response to volume expansion in essential hypertension.


Quantification of cutaneous allergic reactions using 3D optical imaging: A feasibility study.
User-independent quantitative measures of cutaneous allergic reactions can help the physicians manage and evaluate the treatment of cutaneous allergic reactions. In this paper, we present and validate a method to quantify the elevation, volume and area of cutaneous allergic reactions to red tattoos. The skin surface of allergic tattoo reactions was imaged using an optical 3D scanner. The in-house developed analysis tool measured the elevation, volume and area of the lesions, compared to a reference surface. This reference surface was created by 3D interpolation of the skin after manual removal of the lesions. The error of the interpolation tool was validated using a digital arm model. The error of our optical scanner was determined using a 3D printed lesion phantom. The clinical feasibility of the method was tested in 83 lesions in 17 patients. The method showed clear potential to assess skin elevation, volume change and area of an allergic reaction. The validation measurements revealed that the error due to interpolation increases for larger interpolation areas and largely determined the error in the clinical measurements. Lesions with a width ≥4 mm and an elevation ≥0.4 mm could be measured with an error below 26%. Patient measurements showed that lesions up to 600 mm2 could be measured accurately, and elevation and volume changes could be assessed at follow-up. Quantification of cutaneous allergic reactions to red tattoos using 3D optical scanning is feasible and may objectify skin elevation and improve management of the allergic reaction.


Does Written Emotional Disclosure about Stress Improve College Students' Academic Performance? Results from Three Randomized, Controlled Studies.
Several early studies and subsequent reviews suggested that written emotional disclosure (WED)-writing repeatedly about personal stressful experiences-leads to improved academic performance of college students. Yet a critical review of available studies casts some doubt on this, so we conducted three randomized, controlled experiments of the effects of WED versus control writing on grade point average (GPA) of college students. In all three studies, WED writing was implemented effectively-it contained more negative emotion language and generated more negative mood than did control writing. In Study 1, WED did not influence GPA during either the writing semester or subsequent semester among 96 students with headaches. In Study 2, WED had no effect on GPA compared with either control writing or no writing conditions among 124 students with unresolved stress. In Study 3, WED did not influence GPA or retention among 68 academically at-risk ethnic minority students, although secondary analyses suggested some benefits of WED among students who wrote more than once, particularly men. These three studies challenge the belief that WED improves academic performance of college students, and research should examine subgroups of students who might benefit from WED.


The use of patterned dual thermoresponsive surfaces for the collective recovery as co-cultured cell sheets.
Heterotypic cell interactions are critical to achieve and maintain specific functions in many tissues and organs. We have focused on patterned structure surfaces to enable co-culture of heterotypic cells and recovery of patterned co-cultured cell sheets for applications in tissue engineering. Thermoresponsive polymers exhibiting different transition temperatures in water comprise both poly(N-isopropylacrylamide) (PIPAAm) and n-butyl methacrylate (BMA) co-grafted as side chains to PIPAAm main chains. These copolymers were surface-grafted in patterns to obtain patterned dual thermoresponsive cell culture surfaces using electron beam polymerisation method and porous metal masks. On patterned surfaces, site-selective adhesion on and growth of rat primary hepatocytes (HCs) and bovine carotid endothelial cells (ECs) allowed patterned co-culture, exploiting hydrophobic/hydrophilic surface chemistry regulated by culture temperature as the sole variable. At 27 degrees C, seeded HCs adhered exclusively onto hydrophobic, dehydrated P(IPAAm-BMA) co-grafted domains (1-mm laser dot), but not onto neighbouring hydrated PIPAAm domains. Sequentially seeded ECs then adhered exclusively to hydrophobised PIPAAm domains upon increasing culture temperature to 37 degrees C, achieving patterned co-cultures. Reducing culture temperature to 20 degrees C promoted hydration of both polymer-grafted domains, permitting release of the co-cultured, patterned cell monolayers as continuous cell sheets with heterotypic cell interactions. Recovered co-cultured cell sheets can be manipulated, moved and sandwiched with other structures, providing new useful constructs both for basic cell biology research and preparation of tissue-mimicking multi-layer materials through overlaying co-cultured cell sheets.


A model for the passage of the nurohypophysial hormones and their related proteins through the rat neurohypophysis.
The change in the radioactivity of vasopressin-neurophysin in the rat neurohypophysis after an intracisternal injection of [(35)S]cysteine was fitted to several mathematical models. The data fitted best a model in which there is a linear input of radioactive protein into one pool of the neurohypophysis, from which it is either released by an exponential process or transferred to a second pool from which it is released by a second exponential process with a rate constant much lower than the first. This model is compatible with the existence of a ;readily releasable' pool first postulated by Sachs et al. (1967). Data for the change in radioactivity of vasopressin also gave a good fit in this model. Calculation of the rate constants suggested that the first pool represented about 2% of the total hormone.


Evaluating the safety and efficacy of dextromethorphan/quinidine in the treatment of pseudobulbar affect.
Pseudobulbar affect (PBA) is a common manifestation of brain pathology associated with many neurological diseases, including amyotrophic lateral sclerosis, Alzheimer's disease, stroke, multiple sclerosis, Parkinson's disease, and traumatic brain injury. PBA is defined by involuntary and uncontrollable expressed emotion that is exaggerated and inappropriate, and also incongruent with the underlying emotional state. Dextromethorphan/quinidine (DM/Q) is a combination product indicated for the treatment of PBA. The quinidine component of DM/Q inhibits the cytochrome P450 2D6-mediated metabolic conversion of dextromethorphan to its active metabolite dextrorphan, thereby increasing dextromethorphan systemic bioavailability and driving the pharmacology toward that of the parent drug and away from adverse effects of the dextrorphan metabolite. Three published efficacy and safety studies support the use of DM/Q in the treatment of PBA; significant effects were seen on the primary end point, the Center for Neurologic Study-Lability Scale, as well as secondary efficacy end points and quality of life. While concentration-effect relationships appear relatively weak for efficacy parameters, concentrations of DM/Q may have an impact on safety. Some special safety concerns exist with DM/Q, primarily because of the drug interaction and QT prolongation potential of the quinidine component. However, because concentrations of dextrorphan (which is responsible for many of the parent drug's side effects) and quinidine are lower than those observed in clinical practice with these drugs administered alone, some of the perceived safety issues may not be as relevant with this low dose combination product. However, since patients with PBA have a variety of other medical problems and are on numerous other medications, they may not tolerate DM/Q adverse effects, or may be at risk for drug interactions. Some caution is warranted when initiating DM/Q treatment, particularly in patients with underlying risk factors for torsade de pointes and in those receiving medications that may interact with DM/Q.


SOD1 is a synthetic lethal target in PPM1D-mutant leukemia cells.
The DNA damage response is critical for maintaining genome integrity and is commonly disrupted in the development of cancer. PPM1D (protein phosphatase, Mg2+/Mn2+ dependent 1D) is a master negative regulator of the response; gain-of-function mutations and amplifications of PPM1D are found across several human cancers making it a relevant pharmacologic target. Here, we used CRISPR/Cas9 screening to identify synthetic-lethal dependencies of PPM1D, uncovering superoxide dismutase-1 (SOD1) as a potential target for PPM1D-mutant cells. We revealed a dysregulated redox landscape characterized by elevated levels of reactive oxygen species and a compromised response to oxidative stress in PPM1D-mutant cells. Moreover, we observed marked genomic instability in mutant cells, which is exacerbated upon inhibition of SOD1. Altogether, our results demonstrate the protective role of SOD1 against oxidative stress and DNA damage in PPM1D-mutant leukemia cells and highlight a new potential therapeutic strategy against PPM1D-mutant cancers.


Successful interventional treatment of renal insufficiency caused by renal artery pseudoaneurysm with concomitant arteriovenous fistula.
Asymptomatic arteriovenous fistulae and pseudoaneurysms are common after renal biopsy. We present a patient with a single kidney and symptomatic pseudoaneurysm of the renal artery with concomitant arteriovenous fistula as a rare complication following surgical embolectomy. The patient developed renal insufficiency because of a significant degree of vascular steal caused by the fistula. Dialysis was performed for more than 6 months until the fistula was diagnosed. The fistula and the pseudoaneurysm were successfully treated by superselective arterial embolization with metallic coils. Renal function improved within the next 2 weeks and no further dialysis was necessary.


Improved separation method for rat proximal and distal renal tubules.
We modified and improved enzyme digestion and density gradient separation procedures to obtain fractions of proximal and distal renal tubules with high yield and viability. Kidneys from two anesthetized adult Wistar rats were flushed with Krebs-Henseleit buffer (KHB) and then perfused in situ with recirculated KHB containing collagenase and hyaluronidase at 125 mmHg. Cortices were excised, minced, and incubated in KHB containing enzymes for 35 min at 37 degrees C. Dissociated tubules were removed at 10-min intervals, rinsed, and placed in KHB containing 10% calf serum, vitamins, and amino acids at 4 degrees C. Separation was achieved by suspending the tissue in 45% isosmotic Percoll layered over an undiluted Percoll cushion and centrifuging. Proximal tubules sedimented near the cushion. Distal segments were isolated in the uppermost bands of a second 35% Percoll separation. Viability was greater than 95% as measured by lactate dehydrogenase leakage and quantitated by oxygen consumption and ATP content. Basal oxygen consumption was greater than 33 nmol O2 X min-1 X mg protein-1 in all fractions and was stimulated by succinate and inhibited by amiloride and ouabain. Basal ATP content averaged 9.7 nmol/mg ATP. An average 3.3-fold separation for the proximal fraction and 24.5-fold separation for the distal fraction was assessed by the enrichment of six specific enzyme markers, with several of the markers indicating separations up to 32-fold. Isolated tubules also displayed functional responses to parathyroid hormone and vasopressin. Distal, but not proximal, segments demonstrated significantly increased adenosine 3',5'-cyclic monophosphate formation with vasopressin.


[Diagnosis of hemostasis disorders in young children using turbidimetry].
Practical application of turbidimetrical method for the diagnosis of haemostasis disorders in babies with acute intestinal infections (AII) is described. The results of plasma biochemical analysis show that disturbances in balance between coagulation and fibrinolytic systems takes place in patients with this pathology. The correlation between biochemical parameters and clinical characteristics is observed. As follows from our data, the phase of the haemostasis disorders in babies with AII does not depend on etiology of the disease, but indicates its severity.


Versatile morphometric analysis and visualization of the three-dimensional structure of neurons.
The computational properties of a neuron are intimately related to its morphology. However, unlike electrophysiological properties, it is not straightforward to collapse the complexity of the three-dimensional (3D) structure into a small set of measurements accurately describing the structural properties. This strong limitation leads to the fact that many studies involving morphology related questions often rely solely on empirical analysis and qualitative description. It is possible however to acquire hierarchical lists of positions and diameters of points describing the spatial structure of the neuron. While there is a number of both commercially and freely available solutions to import and analyze this data, few are extendable in the sense of providing the possibility to define novel morphometric measurements in an easy to use programming environment. Fewer are capable of performing morphometric analysis where the output is defined over the topology of the neuron, which naturally requires powerful visualization tools. The computer application presented here, Py3DN, is an open-source solution providing novel tools to analyze and visualize 3D data collected with the widely used Neurolucida (MBF) system. It allows the construction of mathematical representations of neuronal topology, detailed visualization and the possibility to define non-standard morphometric analysis on the neuronal structures. Above all, it provides a flexible and extendable environment where new types of analyses can be easily set up allowing a high degree of freedom to formulate and test new hypotheses. The application was developed in Python and uses Blender (open-source software) to produce detailed 3D data representations.


Controlled trials of inositol in psychiatry.
Inositol is a simple polyol precursor in a second messenger system important in the brain. Cerebrospinal fluid inositol has been reported as decreased in depression. A double-blind controlled trial of 12 g daily of inositol in 28 depressed patients for four weeks was performed. Significant overall benefit for inositol compared to placebo was found at week 4 on the Hamilton Depression Scale. No changes were noted in hematology, kidney or liver function. Since many antidepressants are effective in panic disorder, twenty-one patients with panic disorder with or without agoraphobia completed a double-blind, placebo-controlled, four week, random-assignment crossover treatment trial of inositol 12 g per day. Frequency and severity of panic attacks and severity of agoraphobia declined significantly with inositol compared to placebo. Side-effects were minimal. Since serotonin re-uptake inhibitors benefit obsessive compulsive disorder (OCD) and inositol is reported to reverse desensitization of serotonin receptors, thirteen patients with OCD completed a double-blind controlled crossover trial of 18 g inositol or placebo for six weeks each. Inositol significantly reduced scores of OCD symptoms compared with placebo. A controlled double-blind crossover trial of 12 g daily of inositol for a month in twelve anergic schizophrenic patients, did not show any beneficial effects. A double-blind controlled crossover trial of 6 g of inositol daily vs. glucose for one month each was carried out in eleven Alzheimer patients, with on clearly significant therapeutic effects. Antidepressant drugs have been reported to improve attention deficit disorder (ADDH) with hyperactivity symptomatology. We studied oral inositol in children with ADDH in a double-blind, crossover, placebo-controlled manner. Eleven children, mean age 8.9 +/- 3.6 years were enrolled in an eight week trial of inositol or placebo at a dose of 200 mg/kg body weight. Results show a trend for aggravation of the syndrome with myo-inositol as compared to placebo. Recent studies suggest that serotonin re-uptake inhibitors are helpful in at least some symptoms of autism. However a controlled double-blind crossover trial of inositol 200 mg/kg per day showed no benefit in nine children with autism. Cholinergic agonists have been reported to ameliorate electroconvulsive therapy (ECT)-induced memory impairment. Inositol metabolism is involved in the second messenger system for several muscarinic cholinergic receptors. Inositol 6 g daily was given in a crossover-double-blind manner for five days before the fifth or sixth ECT to a series of twelve patients, without effect. These results suggest that inositol has therapeutic effects in the spectrum of illness responsive to serotonin selective re-uptake inhibitors, including depression, panic and OCD, and is not beneficial in schizophrenia, Alzheimer's ADDH, autism or ECT-induced cognitive impairment.


Photobleaching imprinting microscopy: seeing clearer and deeper.
We present a generic sub-diffraction-limited imaging method - photobleaching imprinting microscopy (PIM) - for biological fluorescence imaging. A lateral resolution of 110 nm was measured, more than a twofold improvement over the optical diffraction limit. Unlike other super-resolution imaging techniques, PIM does not require complicated illumination modules or specific fluorescent dyes. PIM is expected to facilitate the conversion of super-resolution imaging into a routine lab tool, making it accessible to a much broader biological research community. Moreover, we show that PIM can increase the image contrast of biological tissue, effectively extending the fundamental depth limit of multi-photon fluorescence microscopy.


[Parasitic infections of coyote, Canis latrans (Carnivora: Canidae) in a Costa Rican National Park and a surrounding agricultural area].
As human populations expand into wild habitats with their pets and livestock, the potential spread of disease to wildlife or vice versa increases. Because, wild and domestic canids may pose as reservoirs or disseminators of infectious diseases (including parasites), coyotes (Canis latrans) may also serve as indicators of ecological health. In Costa Rica, little information exists on coyote parasites, making research necessary to identify potential zoonotic interactions. For this reason, a survey of the coyote parasites was performed in a mixed area of protected woodland and agricultural land, surrounding Irazú Volcano National Park (IVNP) in Cartago, Costa Rica. Over a one-year period, 209 fecal samples were collected directly from the ground. Collection took place on a monthly basis in a trail sectioned into three sub-areas named Irazú (closest to the volcano), potato fields (where potatoes were cultivated), and Prusia (a protected sector of IVNP). Sectioning the trail allowed separate collection and analysis of the samples, where 99 were obtained from Irazú, 11 from potato fields and 99 from Prusia. Using direct examination and mechanical concentration 36.84% positive samples containing at least one helminth were found. The presence of parasites was similar for both woodland areas (33.3% in Prusia and 37.4% in Irazú), but differed from the 63.6% observed in the potato fields. Hookworm parasites (probably Ancylostoma caninum), threadworms (possibly Strongyloides sp.), Toxocara canis, Trichuris sp. and Taenia pisiformis were identified, as well as Hymenolepis diminuta, possible spurious parasite resulting from the ingestion of rodents by coyotes. Seasonal details are discussed, concluding that wet and dry seasons affect presence of parasites. Some remarks are made on the importance of these first findings for Costa Rica, especially considering the systematic way in which the collection of samples was carried out.


CellML and associated tools and techniques.
We have, in the last few years, witnessed the development and availability of an ever increasing number of computer models that describe complex biological structures and processes. The multi-scale and multi-physics nature of these models makes their development particularly challenging, not only from a biological or biophysical viewpoint but also from a mathematical and computational perspective. In addition, the issue of sharing and reusing such models has proved to be particularly problematic, with the published models often lacking information that is required to accurately reproduce the published results. The International Union of Physiological Sciences Physiome Project was launched in 1997 with the aim of tackling the aforementioned issues by providing a framework for the modelling of the human body. As part of this initiative, the specifications of the CellML mark-up language were released in 2001. Now, more than 7 years later, the time has come to assess the situation, in particular with regard to the tools and techniques that are now available to the modelling community. Thus, after introducing CellML, we review and discuss existing editors, validators, online repository, code generators and simulation environments, as well as the CellML Application Program Interface. We also address possible future directions including the need for additional mark-up languages.


Impact of natural disasters on consumer behavior: Case of the 2017 El Niño phenomenon in Peru.
El Niño is an extreme weather event featuring unusual warming of surface waters in the eastern equatorial Pacific Ocean. This phenomenon is characterized by heavy rains and floods that negatively affect the economic activities of the impacted areas. Understanding how this phenomenon influences consumption behavior at different granularity levels is essential for recommending strategies to normalize the situation. With this aim, we performed a multi-scale analysis of data associated with bank transactions involving credit and debit cards. Our findings can be summarized into two main results: Coarse-grained analysis reveals the presence of the El Niño phenomenon and the recovery time in a given territory, while fine-grained analysis demonstrates a change in individuals' purchasing patterns and in merchant relevance as a consequence of the climatic event. The results also indicate that society successfully withstood the natural disaster owing to the economic structure built over time. In this study, we present a new method that may be useful for better characterizing future extreme events.


Transcriptional pausing in a region important for plasmid NR1 replication control.
The results of in vitro single-round transcription experiments indicated that RNA polymerase pauses during transcription of the leader region that precedes the repA1 gene of IncFII plasmid NR1. Transcription initiated at either of the two transcription promoter sites of the repA1 gene, which encodes the essential replication initiation protein of NR1, was observed to pause in this region. Pausing was specifically enhanced by addition of NusA protein, an Escherichia coli transcription accessory factor. Northern blot RNA-DNA hybridization analysis of repA1 mRNA synthesized in vivo revealed RNA species that had lengths equivalent to those of the in vitro-paused intermediates. The steady-state rate of in vivo repA1 mRNA transcription downstream from the pause sites (measured by quantitative hybridization of pulse-labeled RNA to DNA probes complementary to different segments of repA1 mRNA) was not appreciably affected, which suggests that the pause sites do not promote premature termination of transcription. The pause sites were located between the target sequence within the leader region of the mRNA that interacts with a 91-base countertranscript and the beginning of the repA1 coding sequence. Because the countertranscript is an inhibitor of translation of repA1 mRNA, transcriptional pausing in this region may be an important feature of the regulation of RepA1 synthesis, which is the mechanism by which plasmid NR1 controls its replication.


[Methods for studying the fibrinolysis system and its components].
Comparative analysis of the methods of estimation of the state of fibrinolytic system and its components, which are used in clinical practice now is presented in the review. The advantages of the approaches, which are based on the simultaneous determination of several indices of this system were considered. It was shown promising to use these methods in ophthalmology for estimation of both common and local (in liquids and eye tissues) haemostats, as important additional diagnostic and prognostic criteria of development of some ophthalmopathologies.


Hypothermia in acute liver failure.
The development of encephalopathy in patients with acute liver injury defines the occurrence of liver failure. The encephalopathy of acute liver failure is characterized by brain edema which manifests clinically as increased intracranial pressure. Despite the best available medical therapies a significant proportion of patients with acute liver failure die due to brain herniation. The present review explores the experimental and clinical data to define the role of hypothermia as a treatment modality for increased intracranial pressure in patients with acute liver failure.


Gene expression profiling of skeletal muscle in exercise-trained and sedentary rats with inborn high and low VO2max.
The relationship between inborn maximal oxygen uptake (VO(2max)) and skeletal muscle gene expression is unknown. Since low VO(2max) is a strong predictor of cardiovascular mortality, genes related to low VO(2max) might also be involved in cardiovascular disease. To establish the relationship between inborn VO(2max) and gene expression, we performed microarray analysis of the soleus muscle of rats artificially selected for high- and low running capacity (HCR and LCR, respectively). In LCR, a low VO(2max) was accompanied by aggregation of cardiovascular risk factors similar to the metabolic syndrome. Although sedentary HCR were able to maintain a 120% higher running speed at VO(2max) than sedentary LCR, only three transcripts were differentially expressed (FDR <or=0.05) between the groups. Sedentary LCR expressed high levels of a transcript with strong homology to human leucyl-transfer RNA synthetase, of whose overexpression has been associated with a mutation linked to mitochondrial dysfunction. Moreover, we studied exercise-induced alterations in soleus gene expression, since accumulating evidence indicates that long-term endurance training has beneficial effects on the metabolic syndrome. In terms of gene expression, the response to exercise training was more pronounced in HCR than LCR. HCR upregulated several genes associated with lipid metabolism and fatty acid elongation, whereas LCR upregulated only one transcript after exercise training. The results indicate only minor differences in soleus muscle gene expression between sedentary HCR and LCR. However, the inborn level of fitness seems to influence the transcriptional adaption to exercise, as more genes were upregulated after exercise training in HCR than LCR.


Geographical and practical challenges in the implementation of digital health passports for cross-border COVID-19 pandemic management: a narrative review and framework for solutions.
The rapid global spread of infectious diseases, epitomized by the recent COVID-19 pandemic, has highlighted the critical need for effective cross-border pandemic management strategies. Digital health passports (DHPs), which securely store and facilitate the sharing of critical health information, including vaccination records and test results, have emerged as a promising solution to enable safe travel and access to essential services and economic activities during pandemics. However, the implementation of DHPs faces several significant challenges, both related to geographical disparities and practical considerations, necessitating a comprehensive approach for successful global adoption. In this narrative review article, we identify and elaborate on the critical geographical and practical barriers that hinder global adoption and the effective utilization of DHPs. Geographical barriers are complex, encompassing disparities in vaccine access, regulatory inconsistencies, differences across countries in data security and users' privacy policies, challenges related to interoperability and standardization, and inadequacies in technological infrastructure and limited access to digital technologies. Practical challenges include the possibility of vaccine contraindications and breakthrough infections, uncertainties surrounding natural immunity, and limitations of standard tests in assessing infection risk. To address geographical disparities and enhance the functionality and interoperability of DHPs, we propose a framework that emphasizes international collaboration to achieve equitable access to vaccines and testing resources. Furthermore, we recommend international cooperation to establish unified vaccine regulatory frameworks, adopting globally accepted standards for data privacy and protection, implementing interoperability protocols, and taking steps to bridge the digital divide. Addressing practical challenges requires a meticulous approach to assessing individual risk and augmenting DHP implementation with rigorous health screenings and personal infection prevention measures. Collectively, these initiatives contribute to the development of robust and inclusive cross-border pandemic management strategies, ultimately promoting a safer and more interconnected global community in the face of current and future pandemics.


Prostaglandins, leukotrienes, and platelet-activating factor in shock.
Three major lines of evidence support a role of eicosanoids and PAF in shock. Formation of each of the cyclooxygenase metabolites of arachidonate is enhanced at some point during the shock; these metabolites include PGE2, PGF2 alpha, PGI2, and TXA2. Enhanced formation of 5-HETE and the cysteinyl-LTs provides evidence for activation of the 5-lipoxygenase pathway of arachidonate metabolism, and preliminary biochemical evidence suggests that formation of PAF in anaphylactic and endotoxic shock is also enhanced. Second, TXA2, cysteinyl-leukotrienes, and, to an even greater extent, PAF are able to produce shock and death in intact animals. Third, pharmacological studies show that selective antagonists or synthesis inhibitors modify the course of the shock. While any of these lines of evidence may not by itself provide proof for a cause-effect relationship, the data taken together strongly suggest that vasoactive lipids might be involved in fundamental processes in the pathophysiology of shock. However, the role of vasoactive lipids might vary in different shock paradigms, change at various time points during the evolution of the shock, and depend on the species studied. Moreover, while the majority of the reports tend to focus on a specific substance, the metabolism of all of the eicosanoids mentioned, as well as PAF and probably other arachidonate metabolites (e.g. 15-lipoxygenase products such as lipoxins), changes during shock states. This fact probably causes most of the discrepancies in studies using specific antagonists or synthesis inhibitors to modify the state of shock. Thus, while blockade of one mediator might provide some protection, it might not be sufficient to halt or reverse the main course of the pathophysiological process. For example, the increase in vascular permeability, a fundamental phenomenon in trauma, anaphylaxis, or endotoxemia, might be mediated by PAF, LTs, PGs, peptides (e.g. kinins, substance P, CGRP) and amines (e.g. histamine in some species). Attempting to reverse such a complex phenomenon by blocking one specific factor might not be productive unless the specific substance played a key role in generation of the other factors. It seems, however, that while interactions between PGs, LTs, and PAF do occur (31, 32, 70), none of the shock states are crucially dependent on one class of the vasoactive lipids. Therefore, the therapeutic strategy should be based on multiple sites of action, either by drug combinations or multiple actions of a specific drug.(ABSTRACT TRUNCATED AT 400 WORDS)


Dietary selenium and selenoprotein function.
Selenium is a trace mineral and an essential nutrient in the human diet. Selenium is found in soil and water and consequently enters the food chain through the root ways of plants and aquatic organisms. Some areas of the world are low in soil selenium resulting in a selenium deficient population and the appearance of an associated heart disease and bone disorders that can be corrected with dietary selenium. Indeed the requirement for dietary selenium was established by these observations and while selenium deficiency is rare in the West, patients requiring long-term intravenous feedings have also show heart disease associated with a deficiency of selenium in the feeding fluids. Subsequently, it has been established that dietary selenium can improve a wide range of human health conditions even in areas with soil replete in selenium.


Increasing the source/sink ratio in Vitis vinifera (cv Sangiovese) induces extensive transcriptome reprogramming and modifies berry ripening.
Cluster thinning is an agronomic practice in which a proportion of berry clusters are removed from the vine to increase the source/sink ratio and improve the quality of the remaining berries. Until now no transcriptomic data have been reported describing the mechanisms that underlie the agronomic and biochemical effects of thinning. We profiled the transcriptome of Vitis vinifera cv. Sangiovese berries before and after thinning at veraison using a genome-wide microarray representing all grapevine genes listed in the latest V1 gene prediction. Thinning increased the source/sink ratio from 0.6 to 1.2 m2 leaf area per kg of berries and boosted the sugar and anthocyanin content at harvest. Extensive transcriptome remodeling was observed in thinned vines 2 weeks after thinning and at ripening. This included the enhanced modulation of genes that are normally regulated during berry development and the induction of a large set of genes that are not usually expressed. Cluster thinning has a profound effect on several important cellular processes and metabolic pathways including carbohydrate metabolism and the synthesis and transport of secondary products. The integrated agronomic, biochemical and transcriptomic data revealed that the positive impact of cluster thinning on final berry composition reflects a much more complex outcome than simply enhancing the normal ripening process.


Spatiotemporal, optogenetic control of gene expression in organoids.
Organoids derived from stem cells have become an increasingly important tool for studying human development and modeling disease. However, methods are still needed to control and study spatiotemporal patterns of gene expression in organoids. Here we combined optogenetics and gene perturbation technologies to activate or knock-down RNA of target genes in programmable spatiotemporal patterns. To illustrate the usefulness of our approach, we locally activated Sonic Hedgehog (SHH) signaling in an organoid model for human neurodevelopment. Spatial and single-cell transcriptomic analyses showed that this local induction was sufficient to generate stereotypically patterned organoids and revealed new insights into SHH's contribution to gene regulation in neurodevelopment. With this study, we propose optogenetic perturbations in combination with spatial transcriptomics as a powerful technology to reprogram and study cell fates and tissue patterning in organoids.


A novel in vivo cell-wall labeling approach sheds new light on peptidoglycan synthesis in Escherichia coli.
Peptidoglycan synthesis and turnover in relation to cell growth and division has been studied by using a new labeling method. This method involves the incorporation of fluorescently labeled peptidoglycan precursors into the cell wall by means of the cell-wall recycling pathway. We show that Escherichia coli is able to import exogenous added murein tripeptide labeled with N-7-nitro-2,1,3-benzoxadiazol-4-yl (AeK-NBD) into the cytoplasm where it enters the peptidoglycan biosynthesis route, resulting in fluorescent labels specifically located in the cell wall. When wild-type cells were grown in the presence of the fluorescent peptide, peptidoglycan was uniformly labeled in cells undergoing elongation. Cells in the process of division displayed a lack of labeled peptidoglycan at mid-cell. Analysis of labeling patterns in cell division mutants showed that the occurrence of unlabeled peptidoglycan is dependent on the presence of FtsZ, but independent of FtsQ and FtsI. Accumulation of fluorescence at the division sites of a triple amidase mutant (ΔamiABC) revealed that AeK-NBD is incorporated into septal peptidoglycan. AmiC was shown to be involved in the rapid removal of labeled peptidoglycan side chains at division sites in wild-type cells. Because septal localization of AmiC is dependent on FtsQ and FtsI, this points to the presence of another peptidoglycan hydrolase activity directly dependent on FtsZ.


The role of flow rotation in the adult right atrium: a 4D flow cardiovascular magnetic resonance study.
In healthy adults, the right atrium (RA) serves as a reservoir for the systemic flow return from the superior vena cava (SVC) and inferior vena cava (IVC), preparing the two flows to be transferred to the right ventricle (RV) and pulmonary circulation. This study aims to quantify the haemodynamics of the RA and the associated SVC and IVC inflows, which have not been fully understood to date. Eighteen adults with structurally normal hearts underwent 4D flow magnetic resonance imaging. The cardiac cycle was resolved to 20 temporal phases with a spatial resolution of 3 × 3 × 3 mm3. Analysis included objective visualisation of the flow structures in the RA identified by three different vortex identification criteria, kinetic energy (KE), enstrophy and dissipation. KE and helicity flux were also assessed in both caval veins. Vortex identification methods confirmed that in the majority of participants the blood flow from the caval veins filling the RA during ventricular systole is not chaotic, but rather forms an organised pattern of a single coherent forward turning vortex structure. Thirteen participants displayed a single vortex flow structure, four showed multiple vortices and one had a helical flow pattern without a clear vortex structure. A strong positive correlation exists between the flow KE and enstrophy density. This suggests that flow energy in the RA is mainly rotational, part of which is convected by the highly helical SVC and IVC inflows. Multiple vortices tend to be associated with higher dissipation rates in the central RA region due to turbulence. The rotational nature of the flow in the RA maintains KE better than non-rotational flow. RA flow characteristics are highly related to the helicity content in the caval veins, as well as the KE flux intensity. Lower caval helicity or IVC KE flux dominance tends to favour single vortex formation while the opposite tends to lead to multiple vortices or the rare helical flow patterns. Atria lacking single vortex flow are inclined to have a larger energy input from atrial contraction.


Novel electrochemical sensor for the selective recognition of chlorogenic acid.
In this study, a novel sensitive molecularly imprinted electrochemical sensor was constructed for the selective detection of chlorogenic acid (CGA) by deposition of a molecularly imprinted siloxane (MIS) film, prepared by sol-gel process, onto Au bare electrode surface. Initially, a (3-mercaptopropyl)siloxane layer (MSL) was formed on the Au bare surface, followed by a siloxane layer obtained from the acid-catalyzed hydrolysis/condensation of a solution constituted by tetraethoxysilane (TEOS), phenyltriethoxysilane (PTEOS), 3-(aminopropyl)trimethoxysilane (APTMS) and CGA, as a molecular template. After the GCA extraction the MIS imprinted film was electrochemically characterized using differential pulse voltammetry (DPV). The MIS/Au sensor was tested in a solution of the CGA template and other similar molecules. This electrode displayed excellent selectivity towards CGA when compared with structurally similar molecules. Under optimized experimental conditions, the peak current response of the sensor for CGA was linear from 5.0 × 10(-7)mol L(-1) to 1.4 × 10(-5)mol L(-1), and the detection limit was 1.48 × 10(-7)mol L(-1). The MIS/Au sensor was successfully applied for the determination of CGA in coffee and tea samples.


Seven-day Oral Intake of Orthosiphon stamineus Leaves Infusion Exerts Antiadhesive Ex Vivo Activity Against Uropathogenic E. coli in Urine Samples.
Orthosiphon stamineus leaves (Java tea) extract is traditionally used for the treatment of urinary tract infections. According to recent in vitro data, animal infection studies, and transcriptomic investigations, polymethoxylated flavones from Java tea exert antiadhesive activity against uropathogenic Escherichia coli (UPEC). This antiadhesive activity has been shown to reduce bladder and kidney lesion in a mice infection model. As no data on the antivirulent activity of Java tea intake on humans are available, a biomedical study was performed on 20 healthy volunteers who self-administered Orthosiphon infusion (4 × 3 g per day, orally) for 7 days. The herbal material used for the study conformed to the specification of the European Pharmacopoeia, and ultra high-performance liquid chromatography (UHPLC) of the infusion showed rosmarinic acid, caffeic acid, and cichoric acid to be the main compounds aside from polymethoxylated flavones. Rosmarinic acid was quantified in the tea preparations with 243 ± 22 µg/mL, indicating sufficient reproducibility of the preparation of the infusion. Urine samples were obtained during the biomedical study on day 1 (control urine, prior to Java tea intake), 3, 6 and 8. Antiadhesive activity of the urine samples was quantified by flowcytometric assay using pre-treated UPEC NU14 and human T24 bladder cells. Pooled urine samples indicated significant inhibition of bacterial adhesion on day 3, 6 and 8. The urine samples had no influence on the invasion of UPEC into host cells. Bacterial proliferation was slightly reduced after 24 h incubation with the urine samples. Gene expression analysis (qPCR) revealed strong induction of fitness and motility gene fliC and downregulation of hemin uptake system chuT. These data correlate with previously reported datasets from in vitro transcriptomic analysis. Increased bacterial motility was monitored using a motility assay in soft agar with UPEC UTI89. The intake of Java tea had no effect on the concentration of Tamm-Horsfall Protein in the urine samples. The present study explains the antiadhesive and anti-infective effect of the plant extract by triggering UPEC from a sessile lifestyle into a motile bacterial form, with reduced adhesive capacity.


Changes in Taste Perception in Patients with Minor and Major Cognitive Impairment Linked to Alzheimer's Disease Recorded by Gustatory Evoked Potentials.
The need for early diagnosis biomarkers in Alzheimer's disease (AD) is growing. Only few studies have reported gustatory dysfunctions in AD using subjective taste tests. The main purpose of the study was to explore gustatory functions using subjective taste tests and recordings of gustatory evoked potentials (GEPs) for sucrose solution in patients with minor or major cognitive impairment (CI) linked to AD, and to compare them with healthy controls. The secondary objective was to evaluate the relationships between GEPs and the results of cognitive assessments and fasting blood samples. A total of 45 subjects (15 healthy subjects, 15 minor CI patients, 15 major CI patients) were included to compare their gustatory functions and brain activity by recording GEPs in response to a sucrose stimulation. CI groups were combined in second analyses in order to keep a high power in the study. Correlations were made with cognitive scores and hormone levels (ghrelin, leptin, insulin, serotonin). Increased P1 latencies and reduced N1 amplitudes were observed in minor or major patients compared to controls. GEPs were undetectable in 6 major and 4 minor CI patients. Thresholds for sucrose detection were significantly higher in the major CI group than in controls or the minor CI group. No correlation was found with hormone levels. The cortical processing of sensory taste information seems to be altered in patients with minor or major CI linked to AD. This disturbance was identifiable with subjective taste tests only later, at the major CI stage.


Recent Advances in the Development of Non-Invasive Imaging Probes for Cancer Immunotherapy.
The last two decades have witnessed a major revolution in the field of tumor immunology including clinical progress using various immunotherapy strategies. These advances have highlighted the potential for approaches that harness the power of the immune system to fight against cancer. While cancer immunotherapies have shown significant clinical successes, patient responses vary widely due to the complex and heterogeneous nature of tumors and immune responses, calling for reliable biomarkers and therapeutic strategies to maximize the benefits of immunotherapy. Especially, stratifying responding individuals from non-responders during the early stages of treatment could help avoid long-term damage and tailor personalized treatments. In efforts to develop non-invasive means for accurately evaluating and predicting tumor response to immunotherapy, multiple affinity-based agents targeting immune cell markers and checkpoint molecules have been developed and advanced to clinical trials. In addition, researchers have recently turned their attention to substrate and activity-based imaging probes that can provide real-time, functional assessment of immune response to treatment. Here, we highlight some of those recently designed probes that image functional proteases as biomarkers of cancer immunotherapy with a focus on their chemical design and detection modalities and discuss challenges and opportunities for the development of imaging tools utilized in cancer immunotherapy.


Exercise-induced motor improvement after complete spinal cord transection and its relation to expression of brain-derived neurotrophic factor and presynaptic markers.
It has been postulated that exercise-induced activation of brain-derived neurotrophic factor (BDNF) may account for improvement of stepping ability in animals after complete spinal cord transection. As we have shown previously, treadmill locomotor exercise leads to up-regulation of BDNF protein and mRNA in the entire neuronal network of intact spinal cord. The questions arise: (i) how the treadmill locomotor training, supplemented with tail stimulation, affects the expression of molecular correlates of synaptic plasticity in spinal rats, and (ii) if a response is related to BDNF protein level and distribution. We investigated the effect of training in rats spinalized at low thoracic segments on the level and distribution of BDNF immunoreactivity (IR) in ventral quadrants of the lumbar segments, in conjunction with markers of presynaptic terminals, synaptophysin and synaptic zinc. Training improved hindlimb stepping in spinal animals evaluated with modified Basso-Beattie-Bresnahan scale. Grades of spinal trained animals ranged between 5 and 11, whereas those of spinal were between 2 and 4. Functional improvement was associated with changes in presynaptic markers and BDNF distribution. Six weeks after transection, synaptophysin IR was reduced by 18% around the large neurons of lamina IX and training elevated its expression by over 30%. The level of synaptic zinc staining in the ventral horn was unaltered, whereas in ventral funiculi it was decreased by 26% postlesion and tended to normalize after the training. Overall BDNF IR levels in the ventral horn, which were higher by 22% postlesion, were unchanged after the training. However, training modified distribution of BDNF in the processes with its predominance in the longer and thicker ones. It also caused selective up-regulation of BDNF in two classes of cells (soma ranging between 100-400 microm2 and over 1000 microm2) of the ventrolateral and laterodorsal motor nuclei. Our results show that it is not BDNF deficit that determines lack of functional improvement in spinal animals. They indicate selectivity of up-regulation of BDNF in distinct subpopulations of cells in the motor nuclei which leads to changes of innervation targeting motoneurons, tuned up by locomotor activity as indicated by a region-specific increase of presynaptic markers.


Effect of relative amplitude and formant transitions on perception of place of articulation by adult listeners with cochlear implants.
Previous studies have shown that manipulation of a particular frequency region of the consonantal portion of a syllable relative to the amplitude of the same frequency region in an adjacent vowel influences the perception of place of articulation. This manipulation has been called the relative amplitude cue. Earlier studies have examined the effect of relative amplitude and formant transition manipulations upon labeling place of articulation for fricatives and stop consonants in listeners with normal hearing. The current study sought to determine if (a) the relative amplitude cue is used by adult listeners wearing a cochlear implant to label place of articulation, and (b) adult listeners wearing a cochlear implant integrated the relative amplitude and formant transition information differently than listeners with normal hearing. Sixteen listeners participated in the study, 12 with normal hearing and 4 postlingually deafened adults wearing the Nucleus 22 electrode Mini Speech Processor implant with the multipeak processing strategy. The stimuli used were synthetic consonant-vowel (CV) syllables in which relative amplitude and formant transitions were manipulated. The two speech contrasts examined were the voiceless fricative contrast /s/-"sh" and the voiceless stop consonant contrast /p/-/t/. For each contrast, listeners were asked to label the consonant sound in the syllable from the two response alternatives. Results showed that (a) listeners wearing this implant could use relative amplitude to consistently label place of articulation, and (b) listeners with normal hearing integrated the relative amplitude and formant transition information more than listeners wearing a cochlear implant, who weighted the relative amplitude information as much as 13 times that of the transition information.


The sequence and structure of snake gourd (Trichosanthes anguina) seed lectin, a three-chain nontoxic homologue of type II RIPs.
The sequence and structure of snake gourd seed lectin (SGSL), a nontoxic homologue of type II ribosome-inactivating proteins (RIPs), have been determined by mass spectrometry and X-ray crystallography, respectively. As in type II RIPs, the molecule consists of a lectin chain made up of two β-trefoil domains. The catalytic chain, which is connected through a disulfide bridge to the lectin chain in type II RIPs, is cleaved into two in SGSL. However, the integrity of the three-dimensional structure of the catalytic component of the molecule is preserved. This is the first time that a three-chain RIP or RIP homologue has been observed. A thorough examination of the sequence and structure of the protein and of its interactions with the bound methyl-α-galactose indicate that the nontoxicity of SGSL results from a combination of changes in the catalytic and the carbohydrate-binding sites. Detailed analyses of the sequences of type II RIPs of known structure and their homologues with unknown structure provide valuable insights into the evolution of this class of proteins. They also indicate some variability in carbohydrate-binding sites, which appears to contribute to the different levels of toxicity exhibited by lectins from various sources.


Association of HIV status and treatment characteristics with VIA screening outcomes in Malawi: A retrospective analysis.
Although evidence from high-resource settings indicates that women with HIV are at higher risk of acquiring high-risk HPV and developing cervical cancer, data from cervical cancer "screen and treat" programs using visual inspection with acetic acid (VIA) in lower-income countries have found mixed evidence about the association between HIV status and screening outcomes. Moreover, there is limited evidence regarding the effect of HIV-related characteristics (e.g., viral suppression, treatment factors) on screening outcomes in these high HIV burden settings. This study aimed to evaluate the relationship between HIV status, HIV treatment, and viral suppression with cervical cancer screening outcomes. Data from a "screen and treat" program based at a large, free antiretroviral therapy (ART) clinic in Lilongwe, Malawi was retrospectively analyzed to determine rates of abnormal VIA results and suspected cancer, and coverage of same-day treatment. Multivariate logistic regression assessed associations between screening outcomes and HIV status, and among women living with HIV, viremia, ART treatment duration and BMI. Of 1405 women receiving first-time VIA screening between 2017-2019, 13 (0.9%) had suspected cancer and 68 (4.8%) had pre-cancerous lesions, of whom 50 (73.5%) received same-day lesion treatment. There was no significant association found between HIV status and screening outcomes. Among HIV+ women, abnormal VIA was positively associated with viral load ≥ 1000 copies/mL (aOR 3.02, 95% CI: 1.22, 7.49) and negatively associated with ART treatment duration (aOR 0.88 per additional year, 95% CI: 0.80, 0.98). In this population of women living with HIV with high rates of ART coverage and viral suppression, HIV status was not significantly associated with abnormal cervical cancer screening results. We hypothesize that ART treatment and viral suppression may mitigate the elevated risk of cervical cancer for women living with HIV, and we encourage further study on this relationship in high HIV burden settings.


Spatial normalization of 3D brain images using deformable models.
The spatial normalization and registration of tomographic images from different subjects is a major problem in several medical imaging areas, including functional image analysis, morphometrics, and computer-aided neurosurgery. The focus of this article is the development of a computerized methodology for the spatial normalization of 3D images. We propose a technique that is based on geometric deformable models. In particular, we first describe a deformable surface algorithm that finds a mathematical representation of the outer cortical surface. Based on this representation, a procedure for obtaining a map between corresponding regions of the outer cortex in two different images is established. This map is subsequently used to derive a 3D elastic warping transformation, which brings two images into register. The performance of our algorithm is demonstrated on several datasets. In particular, we first test our deformable surface algorithm on MR images. We then register MR images to atlas images. In our third experiment, we apply a procedure for matching distinct cortical features identified through the curvature map of the outer cortex. Finally, we apply our technique to images from elderly individuals with substantial ventricular enlargement, and we show a good registration in the ventricular area and the surrounding brain structures. We present a highly automated methodology for spatial normalization of images, using deformable models. Applications of our methodology include stereotactic normalization of functional and structural images, morphological analysis of the brain, and computer-aided neurosurgery.


Fatigue and metabolism of frog muscle fibers during stimulation and in response to caffeine.
Tension and metabolite concentrations were measured in single frog muscle fibers at 15 degrees C in vitro, in response to electrical stimulation or to immersion in caffeine- or potassium chloride-Ringer. Sarcomere length equaled 2.3 micrometers. Interrupted stimulation for 150 s at 20 Hz or stimulation for 7.5 min at 1 Hz was followed by at least 20 min of fatigue, evidenced by a reduced 200-ms test contraction. Fatigued fibers contracted maximally in potassium chloride- or caffeine-Ringer. They had high lactate and glucose 6-phosphate concentrations and a reduced phosphocreatine (PCr) concentration. Adenosine 5'-triphosphate (ATP) concentration was approximately normal but was markedly reduced by a caffeine contracture. A plot of PCr consumption against the tension-time integral at different stimulation frequencies (25, 35, or 50 Hz) and durations had an intercept of 25.5 nmol PCr/mg protein at time zero and a corrected slope of 0.65 nmol approximately P/mg protein per kg . s . cm-2. Prolonged fatigue is not due to energy exhaustion or to the inability of muscle fibers to consume residual ATP but probably arises from long-lasting interference in excitation-contraction coupling, which can be reversed by KCl- or caffeine-induced release of Ca2+ from intracellular stores.


The opinions of clinical staff regarding neonatal procedural pain in two Norwegian neonatal intensive care units.
Neonates are subjected to numerous painful procedures without sufficient pain management. The aim of this study was to describe the opinions of Norwegian physicians, nurses and nurse assistants who care for neonates, regarding procedural pain in neonates. A replication of a previous questionnaire study was conducted in two Norwegian neonatal intensive care units (NICU's). The questionnaire aimed at evaluating procedure painfulness, the current use of pharmacological agents and comfort measures and the optimal use of both. Ninety members of the clinical staff participated, which is a response rate of 87%. Opinions on how procedural pain is currently and optimally managed differed significantly. Although most respondents rated a majority of the listed procedures as being more than moderately painful, pharmacological agents were rarely used, except for the insertion of a chest tube and endotracheal intubation. Comfort measures were also believed to be underutilized, but not to the same degree as pharmacological agents. Procedural pain in neonates is not sufficiently managed and both pharmacological agents and comfort measures are underutilized, according to clinicians at two Norwegian NICU's.


Laparoscopic repositioning of malfunctioning peritoneal dialysis catheters.
Continuous ambulatory peritoneal dialysis is commonly used for the maintenance of patients with chronic renal failure. A common complication associated with this treatment is outflow obstruction of the catheter. Noninvasive approaches to outflow obstruction such as enemas, infusions of saline, and body position changes are rarely successful in the management of these complications (1). Herein we report a review of our experience with the use of laparoscopy for the evaluation and management of peritoneal dialysis catheters with outflow obstruction.


R-Loop in the replication origin of human mitochondrial DNA is resolved by RecG, a Holliday junction-specific helicase.
Stable RNA-DNA hybrids (R-loops) prime the initiation of replication in Escherichia coli cells. The R-loops are resolved by Escherichia coli RecG protein, a Holliday junction specific helicase. A stable RNA-DNA hybrid formation in the mitochondrial D-loop region is also implicated in priming the replication of mitochondrial DNA. Consistent with this hypothesis, the 3' ends of the mitochondrial R-loop formed by in vitro transcription are located close to the initiation sites of the mitochondrial DNA replication. This mitochondrial R-loop is resolved by RecG in a dose-dependent manner. Since the resolution by RecG requires ATP, the resolution is dependent on the helicase activity of RecG. A linear RNA-DNA heteroduplex is not resolved by RecG, suggesting that RecG specifically recognizes the higher structure of the mitochondrial R-loop. This is the first example that R-loops of an eukaryotic origin is sensitive to a junction-specific helicase. The resolution of the mitochondrial R-loop by RecG suggests that the replication-priming R-loops have a common structural feature recognized by RecG.


RADIUM-226 AND POLONIUM-210 IN LEAF TOBACCO AND TOBACCO SOIL.
Contents of radium-226 and polonium-210 in leaf tobacco and tobacco-growing soils vary with the source. The differences may result from production locality, culture, and curing. The polonium seems to be not entirely derived from the radium; plants probably take it up from the soil or air.


Antimicrobial Peptide Omiganan Enhances Interferon Responses to Endosomal Toll-Like Receptor Ligands in Human Peripheral Blood Mononuclear Cells.
LL-37 is a cationic antimicrobial peptide and the sole human member of cathelicidins. Besides its bactericidal properties, LL-37 is known to have direct immunomodulatory effects, among which enhancement of antiviral responses via endosomal toll-like receptors (TLRs). Omiganan pentahydrochloride is a synthetic cationic peptide in clinical development. Previously, omiganan was primarily known for its direct bactericidal and antifungal properties. We investigated whether omiganan enhances endosomal TLR responses, similar to LL-37. Human peripheral blood mononuclear cells were treated with endosomal TLR3, -7, -8, and -9 ligands in the presence of omiganan. Omiganan enhanced TLR-mediated interferon-α release. Subsequent experiments with TLR9 ligands showed that plasmacytoid dendritic cells were main contributors to omiganan-enhanced IFN production. Based on this type I interferon-enhancing effect, omiganan may qualify as potential treatment modality for virus-driven diseases. The molecular mechanism by which omiganan enhances endosomal TLR responses remains to be elucidated.


Change Detection in Auditory Textures.
Many natural sounds have spectrotemporal signatures only on a statistical level, e.g. wind, fire or rain. While their local structure is highly variable, the spectrotemporal statistics of these auditory textures can be used for recognition. This suggests the existence of a neural representation of these statistics. To explore their encoding, we investigated the detectability of changes in the spectral statistics in relation to the properties of the change. To achieve precise parameter control, we designed a minimal sound texture--a modified cloud of tones--which retains the central property of auditory textures: solely statistical predictability. Listeners had to rapidly detect a change in the frequency marginal probability of the tone cloud occurring at a random time.The size of change as well as the time available to sample the original statistics were found to correlate positively with performance and negatively with reaction time, suggesting the accumulation of noisy evidence. In summary we quantified dynamic aspects of change detection in statistically defined contexts, and found evidence of integration of statistical information.


A comparison of the contractile properties of human myometrium obtained from the upper and lower uterine segments.
This study compared the contractile characteristics of myometrium taken from upper and lower uterine segments. Biopsies were obtained from women undergoing classical caesarean section. Myometrial strips were dissected and mounted in an organ bath, and the contractions were recorded. The cross sectional area of the contractile elements within the strips was measured enabling strips of differing dimensions to be compared. There were no significant differences in the contractile rate and force production produced by myometrium from the upper and lower segments. This study demonstrated that for contractile studies, the use of lower segment is appropriate. The results fail to demonstrate any functional regionality of the human uterus in terms of contractility.


Parallel Algorithms for Image Template Matching on Hypercube SIMD Computers.
This correspondence presents several parallel algorithms for image template matching on an SIMD array processor with a hypercube interconnection network. For an N by N image and an M by M window, the time complexity is reduced from O(N2M2) for the serial algorithm to O(M2/K2 + M * log2 N/K + log2 N * log2 K) for the N2K2-PE system (1 ¿ K ¿ M), or to O(N2M2/L2) for the L2-PE system (L ¿ N). With efficient use of the inter-PE communication network, each PE requires only a small local memory, many unnecessary data transmissions are eliminated, and the time complexity is greatly reduced.


Impact of Synbiotic Therapy on the Quality of Life in Patients with Mild-to-Moderately Active Ulcerative Colitis.
Ulcerative colitis is a chronic inflammatory disease which is affecting the quality of life, workforce and nutrition of patients negatively in the active periods due to its symptoms. This study aims to explore how synbiotic treatment affects the quality of life in patients with mild-to-moderate ulcerative colitis. Forty ulcerative colitis patients with mild-to-moderate activity were enrolled in the study. Patients were split into two even randomized groups as synbiotic (20 patients) and control (20 patients). The synbiotic group received synbiotic therapy and the control group received placebo for 8 weeks. Both groups were compared at the start and the end of therapy according to the quality of life scores. Quality of life was determined using a short form-36 (SF-36) questionnaire. An increase in mean SF-36 scores were found in both groups at the end of the study. Altough this increase was higher in patients received synbiotic therapy, the difference was not statistically significant (p > 0.05). SF-36 scores were higher in patients with mild activity or those in remission in both groups. Synbiotic use provides an increase in the SF-36 score, however, this increase is not statistically significant.


Identification of novel bioactive compounds from Olea europaea by evaluation of chemical compounds in the OliveNet™ library: in silico bioactivity and molecular modelling, and in vitro validation of hERG activity.
As highlighted in the OliveNet™ library, Olea europaea consists of a diverse collection of chemical compounds. We have classified over 600 compounds into 13 main classes and 47 subclasses. Various compounds, including oleuropein and hydroxytyrosol, have been investigated for their potential beneficial effects in multiple human pathologies. However, the vast majority of compounds remain largely unexplored and approximately 50% are currently non-commercially available. Here, we utilise conventional software to characterise the absorption, distribution, metabolism, excretion, and toxicity profile of OliveNet™ compounds. Molecular docking was performed for assessment of P-glycoprotein (P-gp) inhibition and interactions with the human ether-à-go-go-related gene (hERG) channel. Potential hERG ion channel inhibition was calibrated using in vitro patch clamp assays and steered molecular dynamics (SMD) simulations were used to examine membrane permeability of a subset of compounds. Our findings indicated that 313 out of 675 olive compounds were predicted to be absorbed by the gastrointestinal tract. Hydroxytyrosol required the least amount of force to pass through the lipid bilayer compared to elenolic acid diglucoside in SMD simulations. Based on the ADMET and molecular docking data, the hERG inhibitory activities of verbascoside, oleuropein, and hydroxytyrosol were investigated using patch clamp assays and they were identified as non-inhibitors. While the favourable properties of well-known compounds were confirmed, we identified oleuropein aglycone decarboxymethyl dialdehyde acetal form, decarboxymethyl elenolic acid dialdehyde, acetal of decarboxymethyl elenolic acid dialdehyde, methyl malate-β-hydroxytyrosol ester, hydroxytyrosil elenolate, D-(+)-erythro-1-(4-hydroxy-3-methoxy)-214-phenyl-1,2,3-propantriol, (+)-1-acetoxypinoresinol-4″-O-methyl ether, and 3-[1-(hydroxymethyl)-(E)-1-propenyl] glutaric acid as potential candidates for synthesis and further evaluation.


Amount of urinary iodine excretion in residents of Taipei City--a hospital-based study.
The amount of urinary iodine excretion was measured by neutron activation analysis in 87 residents of Taipei City. The mean +/- SE of urinary iodine excretion expressed as ug iodine/g creatinine was 337.2 +/- 33.3 (335.3 +/- 59.5 for males and 338.4 +/- 40.6 for females). For each 10-year age group, from 20-29 to greater than or equal to 70, they were 396.6 +/- 106.1, 396.5 +/- 127.6, 306.5 +/- 116.7, 326.4 +/- 40.4, 338.1 +/- 64.3 and 222.9 +/- 100.6, respectively. Although there were no significant differences in sex and age, the amount of urinary iodine excretion was distributed in a very wide range from 38.6 to 1,526 ug iodine/g creatinine. Only 3% of the subjects excreted less than 50 ug iodine/g creatinine but 42.7% of our subjects excreted more than 300 ug iodine/g creatine. Iodine deficiency is not a real problem in Taipei City. However, about half of the Taipei City residents may take more than the suggested optimum amount of iodine.


Smith-Magenis syndrome: haploinsufficiency of RAI1 results in altered gene regulation in neurological and metabolic pathways.
Smith-Magenis syndrome (SMS) is a complex neurobehavioural disorder characterised by intellectual disability, self-injurious behaviours, sleep disturbance, obesity, and craniofacial and skeletal anomalies. Diagnostic strategies are focused towards identification of a 17p11.2 microdeletion encompassing the gene RAI1 (retinoic acid induced 1) or a mutation of RAI1. Molecular evidence shows that most SMS features are due to RAI1 haploinsufficiency, whereas variability and severity are modified by other genes in the 17p11.2 region for 17p11.2 deletion cases. The functional role of RAI1 is not completely understood, but it is probably a transcription factor acting in several different biological pathways that are dysregulated in SMS. Functional studies based on the hypothesis that RAI1 acts through phenotype-specific pathways involving several downstream genes have shown that RAI1 gene dosage is crucial for normal regulation of circadian rhythm, lipid metabolism and neurotransmitter function. Here, we review the clinical and molecular features of SMS and explore more recent studies supporting possible therapeutic strategies for behavioural management.


Production of human pancreatic ribonuclease in Saccharomyces cerevisiae and Escherichia coli.
Human pancreatic ribonuclease (HP-RNase) has considerable promise as a therapeutic agent. Structure-function analyses of HP-RNase have been impeded by the difficulty of obtaining the enzyme from its host. Here, a gene encoding HP-RNase was designed, synthesized, and inserted into two expression vectors that then direct the production of HP-RNase in Saccharomyces cerevisiae (fused to either an unmodified or a modified a-factor pre-pro segment) or Escherichia coli (fused to the pelB signal sequence). HP-RNase produced in S. cerevisiae was secreted into the medium as an active enzyme, isolable at 0.1-0.2 mg/liter of culture. This isolate was heterogeneous due to extensive glycosylation and incomplete maturation of the pre-pro segment. HP-RNase produced in E. coli with the pET expression system was purified from the insoluble fraction of the cell lysate. Renaturation of the reduced and denatured protein produced active, homogeneous enzyme recoverable at 1 mg/liter of culture. The N terminus of the HP-RNase produced from the bacterial expression system was processed fully in vivo. The yeast system, combined with techniques that allow detection of picograms of ribonuclease activity, offers a sensitive probe for studies of post-translational modification and secretory targeting in eukaryotic cells. The bacterial system enables studies both to reveal new structure-function relationships in ribonucleases and to evaluate the use of HP-RNase as a cytotoxin that is tolerated by the human immune system.


Local perceptions of climate change validated by scientific evidence in the Himalayas.
The Himalayas are assumed to be undergoing rapid climate change, with serious environmental, social and economic consequences for more than two billion people. However, data on the extent of climate change or its impact on the region are meagre. Based on local knowledge, we report perceived changes in climate and consequences of such changes for biodiversity and agriculture. Our analyses are based on 250 household interviews administered in 18 villages, and focused group discussions conducted in 10 additional villages in Darjeeling Hills, West Bengal, India and Ilam district of Nepal. There is a widespread feeling that weather is getting warmer, the water sources are drying up, the onset of summer and monsoon has advanced during last 10 years and there is less snow on mountains than before. Local perceptions of the impact of climate change on biodiversity included early budburst and flowering, new agricultural pests and weeds and appearance of mosquitoes. People at high altitudes appear more sensitive to climate change than those at low altitudes. Most local perceptions conform to scientific data. Local knowledge can be rapidly and efficiently gathered using systematic tools. Such knowledge can allow scientists to test specific hypotheses, and policy makers to design mitigation and adaptation strategies for climate change, especially in an extraordinarily important part of our world that is experiencing considerable change.