SMIM1 is a type II transmembrane phosphoprotein and displays the Vel blood group antigen at its carboxyl-terminus.
Disruption of SMIM1, encoding small integral membrane protein 1, is responsible for the Vel-negative blood type, a rare but clinically-important blood type. However, the exact nature of the Vel antigen and how it is presented by SMIM1 are poorly understood. Using mass spectrometry we found several sites of phosphorylation in the N-terminal region of SMIM1 and we found the initiating methionine of SMIM1 to be acetylated. Flow cytometry analyses of human erythroleukemia cells expressing N- or C-terminally Flag-tagged SMIM1, several point mutants of SMIM1, and a chimeric molecule between Kell and SMIM1 demonstrated that SMIM1 carries the Vel antigen as a type II membrane protein with a predicted C-terminal extracellular domain of only 3-12 amino acids.


Siderophore cross-utilization amongst rhizospheric bacteria and the role of their differential affinities for Fe3+ on growth stimulation under iron-limited conditions.
The majority of bacteria isolated from rhizospheres of Arachis hypogea (Groundnut) and Vigna radiata (Mung bean) predominantly produced catechol-type siderophores except for a few fluorescent pseudomonads that produced hydroxamates in addition to catecholates. The rhizospheric isolates differed in their ability to cross-utilize siderophores produced by other rhizospheric isolates (heterologous); some were highly proficient at utilizing heterologous siderophores, while others were poor cross-utilizers. Isolate G9, which utilized hydroxamate as well as catecholate siderophores, was found to be an efficient siderophore cross-utilizer, while isolates G2 and G6 were poor-utilizers of catecholate and non-utilizers of hydroxamate siderophores. Growth stimulation of two isolates G9 and G6 was seen when grown in the presence of externally supplied heterologous siderophores, which they cross-utilized. The iron-regulated outer membrane protein (IROMP) profiles differed for the most cross-utilizer and the least cross-utilizer strains, but in both the cases no new outer membrane proteins (OMP) were induced in response to the exogenous siderophores supplied. The growth of the organisms in the presence of heterologous siderophores that they failed to cross-utilize led to growth inhibition in the case of isolate G9. This appears to be due to a lower affinity of the siderophore of G9 as compared to the exogenously supplied G6 siderophore. A simple method was devised to measure relative affinities of respective siderophores for iron based on CAS solution decolorization by the siderophore preparations. The effect on the growth of the differential affinities of the siderophores for iron and the interactions of the organisms through cross-utilization is also discussed.


Erratum: Structure of the Complete Dimeric Human GDAP1 Core Domain Provides Insights Into Ligand Binding and Clustering of Disease Mutations.
[This corrects the article DOI: 10.3389/fmolb.2020.631232.].


Renal function in pediatric liver transplantation: a long-term follow-up study.
Renal impairment is a frequent complication after orthotopic liver transplantation (OLT). However, most studies in children use inaccurate renal assessment based on serum creatinine, and long-term follow-up data are lacking. The purpose of this study was to determine incidence, determinants, and progression of long-term chronic renal insufficiency (CRI) in a single-center series of pediatric liver transplant recipients. The true glomerular filtration rate was measured by inulin clearance before and serially after OLT in 69 consecutive patients followed more than 2 years after transplantation. Cumulative incidence of CRI (glomerular filtration rate<60 mL/min/1.73 m2) was determined using a Kaplan-Meier method. A Cox proportional hazard model was performed to identify predictors of CRI. The median age at OLT was 3.2 years. The median follow-up time after OLT was 9.3 years (interquartile range 6.3-11.9). At 10 years post-OLT, the cumulative incidence of CRI was 25%. In a multivariate Cox regression model, arterial hypertension during follow-up as time dependant variable (P=0.03), cyclosporine as primary immunosuppression (P=0.048), and liver diseases with potential renal involvement including inborn errors of metabolism, Alagille syndrome, and hepatic fibrosis (P=0.003) were associated with CRI. Renal function is a major concern long after OLT in children. Renal dysfunction post-OLT may be reduced by optimal control of arterial hypertension, immunosuppression protocols adapted to primary liver disease, and calcineurin inhibitor sparing regimen.


Rodent kinin-forming enzyme systems--I. Purification and characterization of plasma kininogen.
Low molecular weight (LMW) kininogen was purified 70-fold with a 16% yield from fresh rat plasma by DEAE-Sephadex chromatography, ammonium sulfate precipitation, Sephadex G-200 gel filtration, SP-Sephadex chromatography, CM-cellulose chromatography, and Sephadex G-200 gel filtration. Ferguson plots of polyacrylamide gel electrophoretic patterns revealed four bands with relative molecular weights of 64,000, 123,500, 252,436 and 357,900 (ratio of 1:2:4:6). Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis provided a single protein band with a molecular weight of 72,000, suggesting that the four kininogen bands had been caused by the aggregation of a single oligomeric protein. The purified LMW rat kininogen Fraction B (3.9 micrograms bradykinin/mg) was used to elicit an antiserum in the rabbit. Monospecificity of the antiserum was demonstrated by immunoelectrophoresis (Laurell rocket and Grabar methods) and, thus, the homogeneity of the kininogen was also. The purified kininogen (both Fractions A and B) formed kinin with human urinary kallikrein, rat urinary kallikrein and hog pancreatic kallikrein. Murphy-Sturm lymphosarcoma acid protease also formed kinin when incubated with the kininogen at pH 3.0. The isoelectric point for both fractions was at pH 4.3. Amino acid analyses showed the two kininogen fractions to be rich in acidic amino acids and to have a total carbohydrate content of 8.5% consisting of galactose (1.2 to 1.5%), mannose (1.9 to 2.1%), N-acetylglucosamine (4.3 to 5.1%), N-acetylgalactosamine (0.3%), and sialic acid (0.68%).


The ethical use of digital technology in teaching anatomy: A southern African perspective.
The use of 21st Century technology in anatomy teaching and the recent crisis caused by the Coronavirus pandemic has stimulated anatomists to ponder the ethics surrounding the utilisation of digital images from human bodies of known and unknown provenance in teaching. This novel study explores the awareness of southern African anatomy educators regarding the provenance and ethical use of human material in digital resources for E-learning purposes. Anatomy educators (both members and non-members of the Anatomical Society of Southern Africa including postgraduate students in anatomy) located in 15 health sciences facilities in southern Africa were asked to participate in the survey which consisted of an anonymous, cross-sectional, questionnaire conducted on an online research data system, REDCap. While 52% of respondents used E-learning resources sourced from their own departments for teaching, only 58% of these had knowledge of the provenance of the human material used. Of the 72% of respondents using images from external E-learning resources, 64% did not know the provenance of the human material in these resources. Some southern African anatomists considered anonymity as equivalent to informed consent. Regarding the acceptability of unclaimed bodies for online images, 37% of respondents were against the use of these bodies, while 20% indicated that it was acceptable. Personal internal moral conflict was acknowledged regarding the use of material from unclaimed bodies, particularly during crises such as the Coronavirus pandemic when digital resources were limited. Factors such as lack of awareness of provenance, the law in South Africa and using anonymity for consent, influence the ethical behaviour of southern African anatomists. Clear guiding principles would be of value for anatomists globally with respect to consent to the taking and distribution of images, and transparency on the source of the digital images provided in digital texts and online platforms. The establishment of both an oversight and ethics committee at institutions where digital imaging will be used is recommended.


Sequence homology between potato spindle tuber viroid and U3B snRNA.
Sequence homology was found by computer analysis between potato spindle tuber viroid (PSTV) RNA and U3B snRNA of Novikoff hepatoma cells. This homology is colinear in arrangement, extends in length to 81% of the entire U3B snRNA molecule and is involved in the PSTV molecule unique sites which, if depicted in terms of the secondary structure of the circular PSTV molecule, reveal a conspicuous regularity in their location. A strong relation in primary structure between PSTV and U3B snRNA is demonstrated by statistical analysis.


A randomized, controlled trial of tonsillectomy in periodic fever, aphthous stomatitis, pharyngitis, and adenitis syndrome.
We carried out a prospective, randomized, controlled trial to clarify the effect of tonsillectomy on the clinical course of periodic fever, aphthous stomatitis, pharyngitis, and adenitis (PFAPA) syndrome. Twenty-six consecutive children (mean age 4.1 years) with at least 5 PFAPA attacks were recruited from 3 tertiary care pediatric hospitals during 1999-2003 and randomly allocated to tonsillectomy or follow-up alone. They were all followed up with symptom diaries for 12 months. Tonsillectomy was allowed after 6 months in the control group if the attacks recurred. Six months after randomization all 14 children in the tonsillectomy group and 6/12 children in the control group (50%) were free of symptoms (difference 50%, 95% confidence interval 23% to 75%, P < .001). Tonsillectomy was performed on 5/6 of the patients in the control group who still had symptoms after 6 months. The remaining unoperated child in the control group had recurrences of the fever episodes throughout the follow-up, but the symptoms became less severe, and the parents did not choose tonsillectomy. Tonsillectomy appeared to be effective for treating PFAPA syndrome. The fever episodes ceased without any intervention in half of the control subjects. We conclude that although the mechanisms behind this syndrome are unknown, tonsillectomy can be offered as an effective intervention for children with PFAPA.


Early identification and management of graft failure after unrelated cord blood transplantation.
Delayed hematologic recovery is common after unrelated donor umbilical cord blood transplants (UCBT). Clinically it is important to quickly differentiate slow engraftment from graft failure (GF). We report the engraftment data on 110 pediatric UCBT recipients. By day 28, 71 patients achieved an ANC >0.5 x 10(9) per liter, and 6 others died early without recovery. Of the remaining 33 patients who were still neutropenic, 20 eventually attained donor myeloid recovery, 3 died of transplant-related complications or recurrent leukemia and 10 survived without donor-derived hematopoiesis. These patients received a second UCBT 33-95 days after the first transplant, after additional immunosuppression. One patient died early, the remaining nine patients were engrafted; eight demonstrated complete, and one mixed, donor chimerism (with subsequent graft loss). Acute GVHD developed in three, and chronic GVHD in six of the eight engrafted patients. Two patients developed EBV-lymphoproliferative disorder. Infections, especially viral, were common and protracted. Six of 10 patients are alive, 165-1375 (median 1147) days after second UCBT. Chimerism studies correlated with subsequent engraftment course. Any result showing <5% donor cells was associated with irreversible graft loss. In conclusion, early second UCBT after primary GF is a feasible treatment option. Chronic GVHD and viral reactivation are common post transplant.


Implications of neuropeptides in neurological diseases.
Neuropeptides are sufficiently stable to allow valid radioimmunoassay of peptide concentrations in post-mortem human nervous tissue and in human cerebrospinal fluid. Studies have now documented abnormalities of peptide concentrations in degenerative diseases of the brain. Somatostatin concentration is reduced in the hippocampus and neocortex of patients dying with Alzheimer's type dementia. In Huntington's disease, there are reduced concentrations of substance P, met-enkephalin and cholecystokinin in the basal ganglia; in contrast the concentrations of somatostatin and TRH are increased. Immunocytochemical and experimental lesion studies are underway in an attempt to localize the peptide-containing cells affected by these disorders; and the potential role of alterations in neuropeptide function in the pathogenesis, clinical manifestations and therapy of these illnesses is of great interest. Although alterations of CSF peptide concentrations have been reported in a variety of human diseases, interpretation of these results requires knowledge of the origin and disposition of CSF peptides. Future research into the pathology of peptidergic systems will depend on the development of specific peptide antagonists to probe dynamic aspects of peptide function and on the application of the tools of molecular biology, such as specific mRNA assays, to human material.


In vitro susceptibility and molecular analysis of gentamicin-resistant enterococci.
Enterococci with gentamicin MICs of 256 to 1,024 micrograms/mL were evaluated for susceptibility to ampicillin plus gentamicin synergism. Sixteen of eighteen enterococcal isolates were not susceptible to synergistic killing by ampicillin plus gentamicin; 11 possessed aac(6')-aph(2"), and 4 possessed aph(2")-Ic. A gentamicin MIC of 512 or 1,024 micrograms/mL predicted lack of ampicillin/gentamicin synergism, but a gentamicin MIC of 256 micrograms/mL did not. For six enterococcal strains possessing the gentamicin-resistance gene aph(2")-Ic, ampicillin plus dibekacin, ampicillin plus netilmicin, and ampicillin plus amikacin produced synergistic killing in five, three, and two strains, respectively.


African Americans Demonstrate Significantly Lower Serum Alanine Aminotransferase Compared to Non-African Americans.
Normal ranges of serum alanine aminotransferase (ALT) may vary by race. However, results from research studies are contradictory, and many of these studies have included only small numbers of African Americans. We investigated ALT values in patients without evidence of liver disease to determine whether normal ranges differ across race groups. We also evaluated whether a race- and sex-dependent upper limit of normal (ULN) would improve the ability of ALT to predict liver disease compared to the sex-dependent ULN currently in use. We identified ICD9 codes for liver conditions and diabetes in medical records from a sample of 6719 patients. Analysis of variance (ANOVA) was used to assess differences in ALT log-transformed distributions by race. Logistic regression was used to evaluate whether the addition of race to the current sex-dependent ULN improves the ability of ALT to predict liver disease (assessed by area under the receiver operating characteristic curve (AUROC)). Among 1200 patients with BMI 18.5 < 25 and no evidence of liver disease or type 2 diabetes in their medical record, African Americans demonstrated significantly lower ALT (23.47 IU/L; 95% CL 22.87-24.10) than a combined group of Asian American/White/Other patients (25.71 IU/L; 95% CL 24.69-26.77). This difference remained across BMI categories. The race- and sex-dependent model demonstrated significantly better predictive ability than the sex-dependent model (AUROC = 66.6% versus 59.6%, respectively; p < 0.0001). In a large, racially diverse sample, African Americans demonstrated significantly lower ALT compared to non-African Americans; this difference remained as BMI increased. The establishment of race-specific normal ranges for ALT could contribute to better screening and care for African American patients.


Arachidonic acid cascade and platelet-activating factor in the network of eye inflammatory mediators: therapeutic implications in uveitis.
The cellular and biochemical events triggered by uveitis involve a complex array of cells and a heterogeneous network of mediators of intraocular inflammation. Resident cells are activated and inflammatory cells are recruited. Chemical mediators from the arachidonic acid cascade, prostaglandins, hydroxyeicosatetraenoic acids, and leukotrienes, are formed. Several of these metabolites are modulators of cellular functions, but when generated in sustained, excessive amounts, they contribute to enhanced vascular permeability and to the onset of pathophysiological responses. Another very active membrane-derived mediator is platelet-activating factor. This important mediator of immune and inflammatory responses may play a central role in uveitis due to cell priming, since interleukin-1, tumor necrosis factor, and other as yet unidentified mediators are also being generated. The concomitant accumulation of these networks of mediators in various parts of the uveal tract leads to spreading of the intraocular inflammatory response and cellular damage. At both early and late stages of uveitis, the generation of free radicals is also a major contributor to the impairment of function. Free radicals are generated in two distinct sites: in the oxidative burst of recruited white cells and in free radical formation and lipid peroxidation in resident cells. The identification of the cellular events that lead to the accumulation of networks of mediators of inflammation and their effects has important therapeutic implications in uveitis.


A simple two-stage "bailout" technique for the removal of an unyielding ureteric stent.
To describe a simple technique that can be used to manage an unexpected mildly to moderately (<10 mm) encrusted ureteric stent with consummate ease when time, equipment, or experience are unavailable. We present a series of 5 patients with impacted ureteric stents that were difficult to remove owing to presumed encrustation of the upper end. The indwelling time for the stent ranged from 8 to 16 weeks. All 5 patients were managed by insertion of a second ureteric stent alongside the original one. The encrusted stents were successfully retrieved in all 5 patients at a subsequent visit after the insertion of the second stent, without the need for further specialist equipment or expertise. Insertion of a second stent next to an unyielding, encrusted ureteric stent is a safe, simple, and effective technique to aid in its retrieval. We propose that it should be considered in select patients where encrustation is unexpected and an experienced endourologist is unavailable.


Lyme disease: European perspective.
The main clinical features of Lyme borreliosis seem to be the same in Europe and North America; however, the course of erythema migrans is distinct, with multiple erythema migrans and hematogeneous dissemination in early Lyme borreliosis less frequently observed in Europe. Moreover, the skin manifestations borrelial lymphocytoma and acrodermatitis chronica atrophicans are apparently European phenomena. Meningoradiculoneuritis in Lyme neuroborreliosis, with its severe radicular pain, is more prominent in Europe. Similar difficulties exist on both sides of the Atlantic with the serologic diagnosis of Lyme borreliosis.


The critical review of methodologies and approaches to assess the inherent skin sensitization potential (skin allergies) of chemicals. Part I.
To critically review currently available methods, or methods under development (in vivo, in vitro, in silico, etc.) used in the evaluation of skin sensitization potential and their applicability in the derivation of quantitative 'safety thresholds'.


RBFOX2 is critical for maintaining alternative polyadenylation patterns and mitochondrial health in rat myoblasts.
RBFOX2, which has a well-established role in alternative splicing, is linked to heart diseases. However, it is unclear whether RBFOX2 has other roles in RNA processing that can influence gene expression in muscle cells, contributing to heart disease. Here, we employ both 3'-end and nanopore cDNA sequencing to reveal a previously unrecognized role for RBFOX2 in maintaining alternative polyadenylation (APA) signatures in myoblasts. RBFOX2-mediated APA modulates mRNA levels and/or isoform expression of a collection of genes, including contractile and mitochondrial genes. Depletion of RBFOX2 adversely affects mitochondrial health in myoblasts, correlating with disrupted APA of mitochondrial gene Slc25a4. Mechanistically, RBFOX2 regulation of Slc25a4 APA is mediated through consensus RBFOX2 binding motifs near the distal polyadenylation site, enforcing the use of the proximal polyadenylation site. In sum, our results unveil a role for RBFOX2 in fine-tuning expression of mitochondrial and contractile genes via APA in myoblasts relevant to heart diseases.


Chlamydiae in oviducts and uteri of repeat breeder pigs.
Chlamydial infections of the genital organs cause reproductive failure in female pigs, and the uterus is recognized a target tissue for an infection. In contrast, information on the effect of chlamydiae on the porcine oviduct is patchily and inconclusive, although the bacteria are known to cause severe tubal defects in humans and laboratory animals. The aim of this study was to examine the segments ampulla (A), isthmus (I) and utero-tubal junction of the left (n=20) or both (n=22) oviducts, and uteri (U) from 42 culled repeat breeder pigs for chlamydiae using ompA-PCR, partial ompA gene sequencing, immunohistochemistry (IHC) and microscopy of tissue specimens for histopathology. As revealed by PCR, among a total of 26 chlamydia-positive females, 19 were tested positive in one or more segments of one or both oviducts, 14 were found positive in the uterus, and concomitant infections of both organs were observed in 7 of them. Sequencing of 33 PCR products revealed the following chlamydial species: Chlamydophila (Cp.) psittaci (n=18), Cp. abortus (n=2), Chlamydia (C.) suis (n=10), and C. trachomatis (n=3). Immunopositive staining was observed within the surface epithelium (in A, I, U), stromal tissue (in I, U) and muscular layer (in A, I, U). A total of 24 females had inflamed oviductal segments (in A and/or I) and 36 inflamed uteri. However, there was no relationship between histopathology and results of PCR or IHC. In conclusion, chlamydiae were found to infect oviducts and uteri of pigs. Further studies are required to clarify whether chlamydial infection causes specific histopathology and alters tubal function.


Skin scores: a review of clinical scoring systems in dermatology.
Scoring systems are emerging in dermatology to help guide clinical decision-making. This article discusses 4 scoring systems that help prognosticate cases of Stevens-Johnson syndrome/toxic epidermal necrolysis, screen for psoriatic arthritis, differentiate cellulitis from pseudocellulitis, and determine the appropriateness of Mohs micrographic surgery (MMS).


Comorbidity in borderline personality: understanding dynamics in development.
Borderline personality disorder is marked by high levels of comorbidity in both adolescent and adult populations. However, the mechanisms involved in the development of comorbidity in BPD remain unclear. To address this issue, the current paper proposes the use of dynamic mutualism theory as a valuable and underexplored framework for investigating comorbidity in BPD from a developmental perspective. Specifically, we discuss how predictions of dynamic mutualism can be extended to better understand the onset, maintenance, and interplay of BPD symptoms with other forms of psychopathology over time. Moreover, we suggest that mutualistic interactions among internalizing and externalizing features throughout early development may foster the emergence of BPD symptoms in adolescence and beyond. Next, we discuss methodological approaches for testing mutualism and review indirect evidence that supports the role of mutualistic processes in the emergence and maintenance of BPD and its comorbidities. We conclude with methodological cautions and recommendations for future studies.


The fatty acid elongase Bond is essential for Drosophila sex pheromone synthesis and male fertility.
Insects use a spectacular variety of chemical signals to guide their social behaviours. How such chemical diversity arises is a long-standing problem in evolutionary biology. Here we describe the contribution of the fatty acid elongase Bond to both pheromone diversity and male fertility in Drosophila. Genetic manipulation and mass spectrometry analysis reveal that the loss of bond eliminates the male sex pheromone (3R,11Z,19Z)-3-acetoxy-11,19-octacosadien-1-ol (CH503). Unexpectedly, silencing bond expression severely suppresses male fertility and the fertility of conspecific rivals. These deficits are rescued on ectopic expression of bond in the male reproductive system. A comparative analysis across six Drosophila species shows that the gain of a novel transcription initiation site is correlated with bond expression in the ejaculatory bulb, a primary site of male pheromone production. Taken together, these results indicate that modification of cis-regulatory elements and subsequent changes in gene expression pattern is one mechanism by which pheromone diversity arises.


Serotonin Receptor 2B Mediates Mechanical Hyperalgesia by Regulating Transient Receptor Potential Vanilloid 1.
Serotonin [5-hydroxytryptamine (5-HT)], an inflammatory mediator, contributes to inflammatory pain. The presence of multiple 5-HT subtype receptors on peripheral and central nociceptors complicates the role of 5-HT in pain. Previously, we found that 5-HT2B/2C antagonist could block 5-HT-induced mechanical hyperalgesia. However, the types of neurons or circuits underlying this effect remained unsolved. Here, we demonstrate that the Gq/11-phospholipase Cβ-protein kinase Cε (PKCε) pathway mediated by 5-HT2B is involved in 5-HT-induced mechanical hyperalgesia in mice. Administration of a transient receptor potential vanilloid 1 (TRPV1) antagonist inhibited the 5-HT-induced mechanical hyperalgesia. 5-HT injection enhanced 5-HT- and capsaicin-evoked calcium signals specifically in isolectin B4 (IB4)-negative neurons; signals were inhibited by a 5-HT2B/2C antagonist and PKCε blocker. Thus, 5-HT2B mediates 5-HT-induced mechanical hyperalgesia by regulating TRPV1 function.


A review on Alzheimer's disease classification from normal controls and mild cognitive impairment using structural MR images.
Alzheimer's disease (AD) is an irreversible neurodegenerative brain disorder that degrades the memory and cognitive ability in elderly people. The main reason for memory loss and reduction in cognitive ability is the structural changes in the brain that occur due to neuronal loss. These structural changes are most conspicuous in the hippocampus, cortex, and grey matter and can be assessed by using neuroimaging techniques viz. Positron Emission Tomography (PET), structural Magnetic Resonance Imaging (MRI) and functional MRI (fMRI), etc. Out of these neuroimaging techniques, structural MRI has evolved as the best technique as it indicates the best soft tissue contrast and high spatial resolution which is important for AD detection. Currently, the focus of researchers is on predicting the conversion of Mild Cognitive Impairment (MCI) into AD. MCI represents the transition state between expected cognitive changes with normal aging and Alzheimer's disease. Not every MCI patient progresses into Alzheimer's disease. MCI can develop into stable MCI (sMCI, patients are called non-converters) or into progressive MCI (pMCI, patients are diagnosed as MCI converters). This paper discusses the prognosis of MCI to AD conversion and presents a review of structural MRI-based studies for AD detection. AD detection framework includes feature extraction, feature selection, and classification process. This paper reviews the studies for AD detection based on different feature extraction methods and machine learning algorithms for classification. The performance of various feature extraction methods has been compared and it has been observed that the wavelet transform-based feature extraction method would give promising results for AD classification. The present study indicates that researchers are successful in classifying AD from Normal Controls (NrmC) but, it still requires a lot of work to be done for MCI/ NrmC and MCI/AD, which would help in detecting AD at its early stage.


[Lymphoscintigraphy: evaluating lymphatic spread of rectal cancer].
To investigate lymphatic drainage of rectal and lymphatic spread of rectal cancer. Rectal endoscopic lymphoscintigraphy was performed in 37 volunteers and 16 patients. One millilitre dextran marked with (99m)Tc was injected bilaterally into submucosa of the rectum through rectal endoscopy. 1, 2, 3 hours after the injection, rectal lymphoscintigraphy was registered with a computerized gamma camera (TOSHIBA GCA901A). Lymphoscintigraphy in control subjects showed upward epirectal and pararectal nodes. The average number of the nodes was 12 three hours after the injection. Compared with histological node examination in all patients operated upon for rectal cancer, rectal endoscopic lymphoscintigraphy showed a sensitivity of 97.1%, a specificity of 75.0%, an overall accuracy of 87.5%, a false positivity of 25.0%, and false negativity of 8.3%. The method is better for evaluating lymphatic spread of rectal cancer before operation.


Aging of cholinergic synapses in the avian iris. Part I--Biochemical studies.
The present experiments were designed to test acetylcholine (ACh) release from cholinergic nerve endings of the chicken iris during aging. Either a one-step K+ stimulated release scheme or a more strenuous two-step depletion-reloading-release scheme were used. First, we found that independent of the scheme of release, as demonstrated by a lower peak release, slower time to peak and a lower rate of increase, the 3-year-old iris released less 3H-ACh than the 4-month-old iris. In order to analyze this effect of aging, we determined five basic parameters of ACh metabolism, including total acetylcholine (ACh) and choline (Ch) levels, 3H-ACh, 3H-Ch and 3H-phosphorylcholine (PhCh) formed from exogenous 3H-Ch, for each experimental type and for the two ages. There appear to be major differences in the handling of both the total Ch and newly labelled (3H-Ch) pools between the two ages. In addition, the homeostatic control system for Ch levels in the older tissue appears to be less efficient and incapable of handling strenuous release tests. This defect of the older tissue to preserve its Ch pool may, in part, be responsible for the drastic decrease in ACh levels seen in the older iris. Together with the demonstrated reduction in neurotransmitter level and in uptake of Ch, the reduction in release demonstrated here forms a characteristic triade of age-related defects which may have important implications for our understanding of aging processes in cholinergic terminals.


Comparing Direct Oral Anticoagulants and Warfarin for Atrial Fibrillation, Venous Thromboembolism, and Mechanical Heart Valves.
To summarize available data for use of direct oral anticoagulants in nonvalvular atrial fibrillation, venous thromboembolism, and mechanical heart valves including dose-response consistency to offer considerations for pharmacotherapeutic decision-making for oral anticoagulants. A Medline search of English-language studies published between 2000 and March 2015 was conducted to identify pertinent papers using combinations of the following words: apixaban, atrial fibrillation, dabigatran, direct oral anticoagulant, edoxaban, factor IIa inhibitors, factor Xa inhibitors, mechanical heart valves, novel oral anticoagulant, rivaroxaban, venous thromboembolism, and warfarin. Original studies, guidelines, and approved prescribing information were evaluated and included if contributing new or complementary data toward the objective. References for all identified studies were reviewed and entries included if contributory. Randomized controlled trials have established the safety and efficacy of direct oral anticoagulants in atrial fibrillation and venous thromboembolism for most patient groups. Direct oral anticoagulants should not be used in patients with mechanical heart valves until proven safe and effective. There are groups for which questions remain regarding inter-patient dose-response consistency for direct oral anticoagulants. There are postmarketing data suggesting poorer real-world performance of dabigatran relative to clinical trial data. Direct oral anticoagulants offer several advantages over warfarin, and large clinical trial data establish the appropriateness of their use in broad populations. There remain groups for whom the relative benefit and risk of these agents relative to warfarin are uncertain. A patient-specific approach in pharmacotherapeutic decision-making is appropriate.


Introduction of hypothermia for neonates with perinatal asphyxia in the Netherlands and Flanders.
Therapeutic hypothermia was introduced in the Netherlands and Flanders, Belgium, in 2008. Since then, an increasing number of patients has been treated - up to 166 in 2010. Complications and outcome were registered in an online database. The aim of this study was to analyse complications and outcome after implementation. Data were retrieved from an online database to which all centres had contributed. In 3 years, 332 patients were treated. Excluding 24 patients with congenital abnormalities or metabolic disorders, mortality was 31.8%. Of the 210 survivors without congenital malformations, 21 had cerebral palsy, another 19 a developmental delay of more than 3 months at the age of at least 24 months, and 2 had severe hearing loss. The total adverse outcome, combining death and adverse neurodevelopment, in 308 patients without congenital malformations is 45.5%, which is similar to that of the large trials. The introduction of therapeutic hypothermia for neonates with perinatal asphyxia in the Netherlands and Flanders has been rapid and successful, with results similar to findings in the randomised controlled trials.


Sex chromosome aneuploidy detection by noninvasive prenatal testing: helpful or hazardous?
To assess the incidence of sex chromosome aneuploidy (SCA) predicted by noninvasive prenatal testing (NIPT), assess test performance, and compare it with nuchal translucency (NT) screening among patients seen in our prenatal diagnosis center. We identified suspected cases of SCA by reviewing results from all NIPT samples sent from our center to commercial laboratories offering analysis by cell-free DNA between 1 December 2012 and 31 July 2015. Records of pregnancies positive for SCA were reviewed for ultrasound findings, NIPT indications, and karyotype results on maternal, fetal, and postnatal samples. Other SCA cases presenting during this period regardless of NIPT status were identified from genetic counseling and cytogenetics laboratory logbooks. Noninvasive prenatal testing predicted SCA in 18/2851 patients (0.63%). All had diagnostic testing of fetal or newborn samples. No patients terminated pregnancies on the basis of NIPT. NIPT suggested triple X in five cases, two with elevated NT: all were confirmed on karyotype. Two Klinefelter syndrome cases were also accurately predicted by NIPT. NIPT indicated monosomy X in 11 cases. Only one was a true positive. Ten were false positives, with 46, XX found on fetal or newborn karyotype. Maternal karyotype was mosaic (45, X[4], 46, XX[26]) in one case. Over the same time period, four additional cases of 45, X were confirmed on fetal samples, all with cystic hygromas. One of these had had a false negative NIPT result. The remaining patients pursued only direct testing via CVS or amniocentesis. Sex chromosome aneuploidy was frequently suspected on NIPT. False positive rate for monosomy X was surprisingly high (91%). Prediction of other SCA was more accurate. Diagnostic fetal chromosome analysis should be offered after abnormal NIPT or in the presence of cystic hygromas despite normal NIPT. NIPT limitations should be explained in pretest counseling. © 2017 John Wiley & Sons, Ltd.


An Ethics-based Approach to Global Health Research Part 2: Strategies for Overcoming Logistic and Implementation Challenges.
With the growth of global pharmacy partnerships and collaborative research, particularly between high-income countries and low- or middle-income countries, it is necessary to establish best practices for fair and ethical collaboration and research. There is a gap in the pharmacy literature in this regard. Through this commentary, authors will present a pathway for future global health researchers including generating ideas based on mutual needs of the partnership and the community; exploring the importance of regulations including the need to conduct research and partnership projects within the confines of each participant's professional scope of practice, expertise, and licensure; describing the need to develop agreements and the components that should be included in such an agreement; discussing ethical guidelines for research planning, obtaining ethical approval, and planning for adverse events; and illustrating ethical considerations for research implementation with considerations around consent, data collection, linking patients to care after the completion of the study, and dissemination. Global examples, with a pharmacy-specific approach where applicable, within each section highlight the importance of discussion and action around ethics and equity when pursuing collaborative research, recognizing that many of these situations involve difficult decisions.


Human recombinant heat shock protein 70 affects the maturation pathways of dendritic cells in vitro and has an in vivo adjuvant activity.
Heat shock proteins (HSPs) are potent inducers of an antigen-specific immunological response. A role of chaperon of immunogenic peptides and a direct effect on APC activation and function have been described. However, the signal transduction events involved in the activation of human APCs are poorly characterized. We investigated, using human monocyte-derived dendritic cells (DCs), the signal transduction pathways activated by a human recombinant HSP70 (r)HSP70 purified from eukaryotic cells. rHSP70 effectively induced a partial maturation of DCs in vitro and a significant increase in the titers of antigen-specific IgG when used as a vaccine adjuvant in vivo. rHSP70 did not desensitize human DCs to LPS stimulation and retained its adjuvant properties in C3H/HeJ mice, which are LPS-resistant as a result of a mutation in TLR-4, ruling out the potential interference of LPS contamination. Effects on DC maturation and in vivo functions correlate to the ability of rHSP70 to activate IkappaB-alpha/NF-kappaB and ERK1/2 pathways in human DCs. No activation of p38 was induced in the same experimental conditions. Our data suggest that the IkappaB-alpha/NF-kappaB pathway has a critical role in the partial maturation of DCs induced by rHSP70.


Therapy of a Chronically Infected Total Femur Prothesis in a 40-year-old Patient.
We report a case of a patient who was treated in 1999 at the age of 19 years with a proximal femur replacement due to Ewing sarcoma. After several infection-associated exchange procedures, the patient presented with a fistula and chronic infection of a total femur prosthesis in August 2019 at the age of 40 years. Interdisciplinary treatment with a 3-stage exchange procedure and implantation of a silver-coated total femur prosthesis was successfully performed and hip disarticulation could be avoided. After a follow-up of 2 years, the patient showed uneventful wound healing and full weight bearing without crutches.


Antioxidant activity of aryltetralone lignans and derivatives from Virola sebifera (Aubl.).
Aryltetralone lignans bearing methylenedioxy groups (1a-b; 2a-b) were isolated from seeds of Virola sebifera. Their antioxidant activities were evaluated by inhibition of lipid peroxidation as indicated by TBARS and chemiluminescence emission (CL) assays. The lignan 1c, 'having a 2'-hydroxy-4',5'-methylenedioxyphenyl group, was the most active compound with TBARS/CL Q 1/2 values of 0.89 and 0.10 microg/mL, respectively. The catechol derivatives 3 and 4, obtained by demethylenation of lignans 1a and 2a, were of similar activity to 1c, and all were much more effective as antioxidants than alpha-tocopherol.


Isolation of anaerobic and aerobic bacteria from clean surgical wounds: an experimental and clinical study.
A pad imprint method was evaluated for quantitative sampling of anaerobic and aerobic bacteria from clean surgical wounds. The best recovery of bacteria from the pads was obtained when they were treated with a rinsing solution in a 'Stomacher' apparatus. Pads of three different fabrics were compared, but no difference was noted in their efficiency in sampling anaerobic and aerobic bacteria from clean surgical wounds. Sampling of known quantities of Staphylococcus aureus from experimental wounds in rabbits performed with the pad method gave a threefold better recovery than with a swab. The pad method was superior to the wound wash-out method when sampling wounds in hip surgery.


DNA-Protein Interactions Studied Directly Using Single Molecule Fluorescence Imaging of Quantum Dot Tagged Proteins Moving on DNA Tightropes.
Many protein interactions with DNA require specific sequences; however, how these sequences are located remains uncertain. DNA normally appears bundled in solution but, to study DNA-protein interactions, the DNA needs to be elongated. Using fluidics single DNA strands can be efficiently and rapidly elongated between beads immobilized on a microscope slide surface. Such "DNA tightropes" offer a valuable method to study protein search mechanisms. Real-time fluorescence imaging of these interactions provides quantitative descriptions of search mechanism at the single molecule level. In our lab, we use this method to study the complex process of nucleotide excision DNA repair to determine mechanisms of damage detection, lesion removal, and DNA excision.


Reoperation for heterochronic intraductal papillary mucinous neoplasm of the pancreas after bile duct neoplasm resection: A case report.
Intraductal papillary neoplasm of the bile duct (IPNB) and intraductal papillary mucinous neoplasm (IPMN) of the pancreas have similar pathological manifestations. However, they often develop separately and it is rare for both to occur together. Patients presenting with heterochronic IPMN after IPNB are prone to be misdiagnosed with tumor recurrence. A 67-year-old male patient was admitted 8.5 years after IPNB carcinoma and 4 years after the discovery of a pancreatic tumor. A left hepatic bile duct tumor with distal bile duct dilatation was found 8.5 years ago by the computed tomography; therefore, a left hepatectomy was performed. The postoperative pathological diagnosis was malignant IPNB with negative cutting edge and pathological stage T1N0M0. Magnetic resonance imaging 4 years ago showed cystic lesions in the pancreatic head with pancreatic duct dilatation, and carcinoembryonic antigen continued to increase. Positron emission tomography showed a maximum standard uptake value of 11.8 in the soft tissue mass in the pancreatic head, and a malignant tumor was considered. Radical pancreatoduodenectomy was performed. Postoperative pathological diagnosis was pancreatic head IPMN with negative cutting edge, pancreaticobiliary type, stage T3N0M0. He was discharged 15 d after the operation. Follow-up for 6 mo showed no tumor recurrence, and quality of life was good. IPNB and IPMN are precancerous lesions with similar pathological characteristics and require active surgery and long-term follow-up.


Transpedicular vertebroplasty after intravertebral cavity formation versus conservative treatment for osteoporotic burst fractures.
There has been no study regarding the comparison between vertebroplasty and conservative treatment for osteoporotic burst fracture. To compare the results of vertebroplasty after intravertebral cavity formation with that of conservative treatment alone. A case-control study. The vertebroplasty group included 40 consecutive patients with primary osteoporotic burst fracture who underwent vertebroplasty using calcium phosphate cement, and the control group was made up of 40 patients given conservative treatment alone who were matched for age, gender, the injury level, and the type of fracture. Two groups of patients who had no neural deficit were prospectively examined. Clinical and radiological outcomes of these two groups were compared blindly. The authors do not report any conflict of interest in this study. Outcome measures included visual analog scale (VAS) of the back pain, analgesic requirements, and mobility; in a lateral radiograph, the deformity index was measured and expressed as the ratio of the vertebral body (VB) height (sum of measurements at anterior, middle, and posterior regions of VB) to the longitudinal diameter of VB. Anteroposterior height comparison was expressed as the percentage of VB height at the anterior region compared with the height at the posterior region. Each recovery rate was calculated as a percentage by the formula: (value at the examination-value before treatment)/value before treatment × 100. The duration of follow-up was more than 12 months (mean; 22.5 months). The mean VAS at 12 months after injury was 2.17 cm in the conservative group and 0.61 cm in the vertebroplasty group (p=.0002). The mean duration of analgesic medication required was 157.2 days in the conservative group and 21.3 days in the vertebroplasty group (p=.0048). The mean deformity index at 12 months after injury was 1.66 in the vertebroplasty group and 1.38 in the control group, and the mean recovery rate was +7.3% and -18.4%, respectively (p<.0001). Anteroposterior height comparison at 12 months after injury was 49.1% in the conservative group and 71.2% in the vertebroplasty group, and the mean recovery rate was -26.4% and +30.1%, respectively (p<.0001). One fracture in the vertebroplasty group showed delayed union until 6 months after injury. In the control group, there were six delayed union including four pseudoarthroses at over 6 months after injury that caused leg pain and were treated by conservative therapy. We conclude that vertebroplasty after intravertebral cavity formation provided a better clinical and radiological result than conservative treatment for osteoporotic burst fracture.


Transcriptional activity in in vitro produced bovine two- and four-cell embryos.
The objectives of this study on in vitro produced bovine two- and four-cell embryos were (1) to investigate the uptake of 3H-uridine through the plasma membrane, (2) to characterize the pattern of RNA synthesis during the second cell cycle, and (3) to measure the incorporation of 3H-uridine into de novo synthesized RNA. A total of 200 embryos were incubated with 3H-uridine for 15, 30, 60 (two- and four-cell embryos), 120 (four-cell embryos), 180 (two-cell embryos), and 240 min (two- and four-cell embryos), respectively. 3H-uridine uptake reached a maximum by 30 min in two-cell embryos, whereas four-cell embryos reached a maximum at 120 min. A total of 440 two-cell embryos were isolated 27-33 hr postinsemination (hpi), and 90 of these were incubated for 10 hr with 3H-uridine (200 microCi/ml). The remainder were incubated with 3H-uridine for 3 hr starting at 0-3 (n = 54), 3-6 (n = 75), 6-9 (n = 77), or 9-12 (n = 77) hr after cleavage to the two-cell stage. Control two-cell embryos (n = 67) were incubated with 3H-uridine supplemented with 5 mg/ml of unlabelled uridine for 10 hr (inhibition control), or they were incubated with 3H-uridine for 10 hr and RNase treated (100 micrograms/ml) post fixation (RNase control). Subsequently, the embryos were processed for autoradiography. The long-term incubation revealed transcription (autoradiographically labelled nuclei) in a total of 77% of the two- and four-cell embryos. No transcription was observed in any of the 3 hr incubation groups. The RNase control embryos lacked labelling of the nuclei, whereas the inhibition control embryos only showed markedly reduced labelling. Finally, total RNA extraction was performed on a total of 336 two-cell embryos that were incubated with 3H-uridine or 3H-uridine supplemented with unlabelled uridine for 2, 5, or 10 hr. It was possible to detect an increasing amount of labelled RNA after the 2, 5, and 10 hr incubation periods, and it was possible to inhibit this incorporation competitively. Together the data demonstrate a low level of transcription during the second cell cycle without a well-defined transcriptional peak.


Drying kinetics and quality dynamics of ultrasound-assisted dried selenium-enriched germinated black rice.
Black rice is a functional food due to its higher protein, fiber, iron, antioxidant compounds, and other health benefits than traditional rice. The ultrasonic (US) pretreatments (10, 20, and 50 min) followed by hot-air drying (50, 60, and 70 °C) were applied to study the drying kinetics, mathematical modeling, thermodynamics, microstructure, bioactive profile, volatile compounds and to lock the nutritional composition of selenium-enriched germinated black rice (SeGBR). Ultrasonic-treated samples exhibited a 20.5% reduced drying time than control ones. The Hii model accurately describes the drying kinetics of SeGBR with the highest R2 (>0.997 to 1.00) among the fifteen studied models. The activation energy values in US-SeGBR varied from 3.97 to 13.90 kJ/mol, while the specific energy consumption ranged from 6.45 to 12.32 kWh/kg, which was lower than untreated. The obtained thermodynamic attributes of dried black rice revealed that the process was endothermic and non-spontaneous. Gallic acid, kaempferol, and cyanidin 3-glucoside were present in high concentrations in phenolics, flavonoids, and anthocyanins, respectively. The HS-SPME-GC-MS investigation detected and quantified 55 volatile compounds. The US-treated SeGBR had more volatile compounds, which may stimulate the release of more flavorful substances. The scanning electronic micrograph shows that the US-treated samples absorbed high water through several micro-cavities. Selenium concentration was significantly higher in US-treated samples at 50 °C than in control samples. In conclusion, ultrasound-assisted hot-air drying accelerated drying and improved SeGBR quality, which is crucial for the food industry and global promotion of this healthiest rice variety.


Going beyond the evidence: abstract laws and preschoolers' responses to anomalous data.
Given minimal evidence about novel objects, children might learn only relationships among the specific entities, or they might make a more abstract inference, positing classes of entities and the relations that hold among those classes. Here we show that preschoolers (mean: 57 months) can use sparse data about perceptually unique objects to infer abstract physical causal laws. These newly inferred abstract laws were robust to potentially anomalous evidence; in the face of apparent counter-evidence, children (correctly) posited the existence of an unobserved object rather than revise the abstract laws. This suggests that children's ability to learn robust, abstract principles does not depend on extensive prior experience but can occur rapidly, on-line, and in tandem with inferences about specific relations.


Association between Intracanal Medicament Radiopacity and Streak Artifact Production Using Cone-beam Computed Tomography: A Laboratory Study.
This study aimed to calculate the correlation between the radiopacity levels of various intracanal medicaments and radiolucent streak formation using cone-beam computed tomography (CBCT). Seven commercially-available intracanal medicaments were tested, which contained different amounts of radiopacifier [Consepsis, Ca(OH)2, UltraCal XS, Calmix, Odontopaste, Odontocide, and Diapex Plus]. Their radiopacity levels were measured according to the International Organization for Standardization 13116 testing standards (mmAl). Subsequently, the medicaments were placed in 3 canals of radiopaque artificial printed maxillary molars (n = 15 roots per medicament), leaving the second mesiobuccal canal empty. CBCT imaging was carried out using an Orthophos SL 3-dimensional scanner under recommended manufacturer exposure settings. Radiopaque streak formation was assessed by a calibrated examiner using a previously published grading system (0-3). The Kruskal-Wallis and Mann-Whitney U tests with and without Bonferroni correction were used to compare radiopacity levels and radiopaque streak scores for the medicaments. Their relationship was assessed using the Pearson correlation coefficient. The level of significance was set as (α = 0.05). Diapex plus presented with the highest radiopacity levels (4.98 ± 0.01) and radiopaque streak scores [middle third (2.8 ± 0.18); apical third (2.73 ± 0.43)]; which was similar to UltraCal XS's radiopaque streak scores [middle third (2.8 ± 0.92); apical third (2.73 ± 0.77)]. Consepsis had the lowest radiopacity levels (0.12 ± 0.05), followed by Odontocide (0.60 ± 0.05). Consepsis and Ca(OH)2 were scored 0 for artifacts in all roots at all levels. A high positive correlation (R = 0.95) was found between radiopacity and streak formation. The radiopacity of intracanal medicaments varies and strongly correlates with the formation of radiolucent streak artifacts during CBCT.


Chiral biscavitand propellers: synthesis, conformations and multiple guest binding.
[structure: see text]. The synthesis and properties of molecules containing two cavitand bowls, with interbowl linkages comprising one carbon atom at various oxidation levels, are reported. A carbonyl-bridged biscavitand binds three CH2Cl2 molecules in the solid state through a helical conformation. Evidence for the same chiral conformation in the solution phase is presented.


The influence of foehn winds on the incidence of severe injuries in southern Bavaria - an analysis of the TraumaRegister DGU®.
Foehn describes a wind which occurs in areas with close proximity to mountains. The presence of foehn wind is associated with worsening health conditions. This study analyzes the correlation between a foehn typical circulation and the incidence for suffering a severe trauma. This is a retrospective, multicentre observational register study. The years from 2013 to 2016 were analyzed for the presence of foehn winds. A logistic regression analysis with the number of daily admitted trauma patients as the primary target value was performed in dependence of foehn winds. Southern Bavaria is a typical foehn wind region. Individuals were treated in 37 hospitals of Southern Bavaria which participate in the TraumaRegister DGU®, an international register that includes all severe trauma patients, mainly in Germany. We analyzed patients with an Injury Severity Score (ISS) of at least nine with admission to intensive care units or prior death in the emergency room. 6215 patients were enrolled in this study. A foehn-typical circulation was present on 65 days (4.5%). 301 patients (5%) suffered a trauma with an ISS ≥ 9 on a foehn day. The mean ISS was 20.2 (9-75). On average, 4.3 patients (0-15 patients) were admitted on a daily basis due to a severe trauma. The multivariate regression analysis revealed a daily increase of 0.87 individuals (p = 0.004; 95% CI 0.23-1.47) on foehn days. During spring 1.07 patients (p = < 0.001; 95% CI 0.72-1.42), in summer 1.98 patients (p = < 0.001; 95% CI 1.63-2.32), in fall 0.63 (p = < 0.001; 95% CI 0.28-0.97) and on Saturdays, 0.59 patients (p = < 0.001; 95% CI 0.24-0.93) were additionally admitted due to severe trauma. Foehn winds are significantly associated with severe trauma in trauma centers of the TraumaNetzwerk DGU®.


Isolation of aggresomes and other large aggregates.
Upon permanent stresses and in various diseases, small protein aggregates may accumulate in cells and cause toxicity. A recently discovered protective system transports these aggregates to the centrosome location via microtubules, to form a large agglomerate of aggregates called the aggresomes. Here, we describe a newly developed method for isolating aggresomes. This principle can also be used for purification of other large structures and even organelles.


Interaction between aldosterone and renomedullary prostaglandins. Competitive action between aspirin and spironolactone.
Aldosterone injected i.m. decreased the release of renomedullary PGEs and the index (urinary Na/K ratio) in conscious normotensive intact and adrenalectomized rats. Coadministration of spironolactone increased the release of PGEs as well as the index (urinary Na/K ratio). The effect of spironolactone was partly inhibited by aspirin injected in a ratio 5:1 (aspirin:spironolactone), and effect which could be reversed by the infusion of a synthetic prostaglandin (PGA2) in a subhypotensive dose.


Long term visuo-vestibular mismatch in freely behaving mice differentially affects gaze stabilizing reflexes.
The vestibulo-ocular reflex (VOR) and the optokinetic reflex (OKR) work synergistically to stabilize gaze in response to head movements. We previously demonstrated that a 14-day visuo-vestibular mismatch (VVM) protocol applied in freely behaving mice decreased the VOR gain. Here, we show for the first time that the OKR gain is also reduced and report on the recovery dynamics of both VOR and OKR after the end of the VVM protocol. Using sinusoidally-modulated stimulations, the decreases in VOR and OKR were found to be frequency-selective with larger reductions for frequencies < 0.5 Hz. Constant-velocity OKR stimulation tests demonstrated that the persistent components of the OKR were not modified while the transient, initial responses were. To identify the signals driving VOR and OKR reductions, we compared the responses of mice exposed to a high-contrast and no-contrast VVM. Despite being more robust in the high-contrast conditions, reductions were largely comparable and recovered with a similar time course. An analysis that directly compared VOR and OKR responses revealed that, alterations in the VOR were of significantly larger amplitude with significantly slower dynamics of recovery. Our findings are evidence for a frequency-selective influence of visual signals in the tuning of gaze stabilizing reflexes in normal mice.


Achieving broad substrate specificity in damage recognition by binding accessible nondamaged DNA.
The structure of the Rad4/Rad23 protein, shown in a recent issue of Nature (Min and Pavletich, 2007), reveals how structurally diverse lesions are recognized in eukaryotic nucleotide excision repair: by probing for accessible nondamaged DNA opposite the lesion.


The Effect of Opioid Stewardship Interventions on Key Outcomes: A Systematic Review.
We sought to identify potential patient safety practices to reduce high-risk opioid prescribing. We conducted a systematic review of the literature to identify opioid stewardship (OS) strategies implemented in primary care and other settings. Included studies evaluated an OS strategy or a multicomponent OS initiative to address potential harms of opioids and used experimental or quasi-experimental designs. We identified 14 studies and 1 systematic review that met inclusion criteria. Most studies examined multicomponent OS interventions, which often consisted of guideline-recommended clinical interventions or care processes (e.g., use urine drug screening, check Prescription Drug Monitoring Program), as well as implementation strategies (e.g., dashboards, audit and feedback). Most studies examined the effect of OS interventions on reducing the potential risks of opioids with judicious prescribing and guideline-concordant care (e.g., reduce inappropriate high opioid dosages, avoid co-prescribing opioids and benzodiazepines, use urine drug screening, treatment agreements). The strength of the evidence is low to moderate that OS efforts decrease numbers of opioid prescriptions, proportion of patients on long-term opioids, or days' supply. The strength of the evidence for OS initiatives producing significant reductions in opioid dosages was moderate. Future research is needed on the effectiveness of OS interventions, particularly studies with experimental designs and in diverse settings within the health care system.


Dexmedetomidine can stabilize hemodynamics and spare anesthetics before cardiopulmonary bypass.
We previously confirmed the effectiveness of dexmedetomidine (DEX) for stabilizing hemodynamics as well as sparing anesthetics during anesthetic induction in patients undergoing cardiac surgery (Kunisawa et al. in J Clin Anesth 21:194-199, 1). In this study, we investigated whether these effects of DEX continue until the start of cardiopulmonary bypass (CPB). Twenty-two patients with mild to moderate cardiovascular disease were randomized into two groups [DF2 group: DEX dose of 0.7 μg/kg/h after initial dose and effect-site concentration (ESC) of fentanyl of 2 ng/ml; PF4 group: saline and ESC of fentanyl of 4 ng/ml]. Propofol was administered for anesthetic induction and maintenance. Hemodynamics, cardiovascular drugs, ESC of propofol, and cardiovascular responses to skin incision (SI) and sternotomy (St) were measured or calculated. Blood pressure (BP) at the pre-/post-SI periods was higher in the DEX group (137 ± 17/140 ± 16 mmHg) than in the placebo group (85 ± 9/109 ± 24 mmHg). Percent increases in cardiovascular response to SI or St were lower in the DEX group than in the placebo group (for example, 1.9 ± 2.2 vs. 27.4 ± 19.9% in systolic BP due to SI). ESCs of propofol at SI and St in the DEX group were lower than those in the placebo group. DEX combined with 2 ng/ml fentanyl before CPB can suppress the decrease in blood pressure at the pre- and post-SI periods, can blunt the cardiovascular responses to SI and St, and can spare the required ESC of propofol despite fentanyl concentration, which was half of that in the placebo group.


Kinetic characterization of inhibition of gamma-aminobutyric acid uptake into cultured neurons and astrocytes by 4,4-diphenyl-3-butenyl derivatives of nipecotic acid and guvacine.
The effects of N-(4,4-diphenyl-3-butenyl) derivatives of nipecotic acid (SKF-89976-A and SKF-100844-A) and guvacine (SKF-100330-A) on neuronal and astroglial gamma-aminobutyric acid (GABA) uptake were investigated. In addition, the uptake of SKF-89976-A was studied using the tritiated compound. All of the compounds were found to be competitive inhibitors of GABA uptake irrespective of the cell type, with Ki values similar to or lower than those of the parent amino acids. Moreover, none of the compounds exhibited selectivity with regard to inhibition of neuronal and glial GABA uptake. In spite of the competitive nature of SKF-89976-A, the compound was not transported by the GABA carriers in the two cell types, because no saturable uptake could be demonstrated.


Fluoroolefins as amide bond mimics in dipeptidyl peptidase IV inhibitors.
The synthesis, selectivity, rat pharmacokinetic profile, and drug metabolism profiles of a series of potent fluoroolefin-derived DPP-4 inhibitors (4) are reported. A radiolabeled fluoroolefin 33 was shown to possess a high propensity to form reactive metabolites, thus revealing a potential liability for this class of DPP-4 inhibitors.


CT staging and surveillance of the thorax in patients with newly diagnosed and recurrent squamous cell carcinoma of the head and neck: is it necessary?
The detection of distant metastases or synchronous primary tumours at initial presentation, or at recurrence in patients with head and neck squamous cell carcinoma (HNSCC), frequently alters the selection of therapy in these patients. A number of series report appreciably high rates for these lesions. This study evaluated 108 computed tomography (CT) scans and chest radiographs (CXR) of the thorax, in 80 patients presenting with HNSCC over a 4 year period. There were three clinical settings; (a) at original diagnosis n = 61, (b) clinical evidence of local/regional recurrence n = 19 (c) suspicion of recurrence due to neck symptomatology n = 28. CT thorax detected two out of 61 (3%) distant metastases at the initial diagnosis stage (both were either stage III or IV) and one out of 19 (5%) patients evaluated at the time of loco/regional recurrence. CXR failed to reveal evidence of pulmonary metastases in the two patients at initial diagnosis stage, but correctly identified pulmonary metastases in the loco-regional recurrence patient. There was no thoracic malignancy detected in the surveillance CT scans, and no synchronous second primary tumour detected during the study. CT is known to be more sensitive than conventional CXR in detecting thoracic pathology in HNSCC patients, however, we feel CT is of limited value in stage I or II disease. We no longer carry out routine staging CT scans of the thorax in patients presenting with stage I or II HNSCC, or with neck symptomatology with no clinical evidence of recurrence.


Missense mutation Leu72Pro located on the carboxyl terminal amphipathic helix of apolipoprotein C-II causes familial chylomicronemia syndrome.
Chylomicronemia syndrome can be caused by 2 autosomal recessive disorders - lipoprotein lipase (LPL) deficiency and apolipoprotein C-II (apo C-II) deficiency. We described 2 siblings with chylomicronemia syndrome of a consanguineous family. To determine the molecular basis of chylomicronemia syndrome in this family, we performed direct DNA sequencing of the LPL and APOC2 genes of the proband. A novel homozygous mutation, Leu72Pro, in the APOC2 gene was found in both siblings whereas their parents were carriers. No LPL mutations were detected in the siblings. Apo C-II contains 3 amphipathic alpha helices; the C-terminal alpha helix is composed of residues 64 to 74. Substitution of residue 72 from a helix former leucine to a helix breaker, proline, is predicted to change the secondary structure of the C-terminal helix and subsequently alter the interaction between apo C-II and LPL. To our knowledge, Leu72Pro is the first missense mutation identified in the C-terminal of apo C-II. The result is consistent with the current biochemical and structural findings that the C-terminal helix of apo C-II is important for activation of LPL.


EPR-kinetic isotope effect study of the mechanism of radical-mediated dehydrogenation of an alcohol by the radical SAM enzyme DesII.
The radical S-adenosyl-L-methionine enzyme DesII from Streptomyces venezuelae is able to oxidize the C3 hydroxyl group of TDP-D-quinovose to the corresponding ketone via an α-hydroxyalkyl radical intermediate. It is unknown whether electron transfer from the radical intermediate precedes or follows its deprotonation, and answering this question would offer considerable insight into the mechanism by which the small but important class of radical-mediated alcohol dehydrogenases operate. This question can be addressed by measuring steady-state kinetic isotope effects (KIEs); however, their interpretation is obfuscated by the degree to which the steps of interest limit catalysis. To circumvent this problem, we measured the solvent deuterium KIE on the saturating steady-state concentration of the radical intermediate using electron paramagnetic resonance spectroscopy. The resulting value, 0.22 ± 0.03, when combined with the solvent deuterium KIE on the maximum rate of turnover (V) of 1.8 ± 0.2, yielded a KIE of 8 ± 2 on the net rate constant specifically associated with the α-hydroxyalkyl radical intermediate. This result implies that electron transfer from the radical intermediate does not precede deprotonation. Further analysis of these isotope effects, along with the pH dependence of the steady-state kinetic parameters, likewise suggests that DesII must be in the correct protonation state for initial generation of the α-hydroxyalkyl radical. In addition to providing unique mechanistic insights, this work introduces a unique approach to investigating enzymatic reactions using KIEs.


Predominant low-molecular-weight proteins in isolated brain capillaries are histones.
Blood-brain barrier (BBB) function is endowed by the expression of unique proteins within the brain capillary endothelium. In the absence of knowing the function of BBB-specific proteins, one strategy for identification of these proteins is the purification and amino acid sequencing of proteins within the brain capillary that are not found in other cells. Earlier studies have shown that a 16-18K triplet of low-molecular-weight proteins in isolated brain capillaries is not found in either erythrocytes or in capillary-free preparations of synaptosomal proteins. Therefore, the present studies describe the purification of the 16-18K triplet of proteins as well as a 14K protein in isolated brain capillaries using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and C4 reverse-phase HPLC. Amino acid sequencing of the N-terminus of the 14K, 17K, and 18K proteins and of two tryptic peptides of the 16K protein showed that these proteins are alpha-globin, histone 2B, histone 3, and histone 2A, respectively. SDS-PAGE of subcellular fractions of bovine brain capillaries demonstrated that the 16-18K triplet of histone proteins migrated in the nuclear fraction. In addition, a 34K doublet and a 200K protein were localized in the nuclear pellet. Therefore, the present studies demonstrate that the predominant 14-18K proteins seen on SDS-PAGE of isolated brain capillaries are known proteins and provide a general scheme for purification of brain capillary proteins isolated following SDS-PAGE.


The coalescent in two colonies with symmetric migration.
Kingman's coalescent process is extended to two colonies with symmetric migration. The mean waiting time until a sample of genes taken from two colonies coalesces to a common ancestor is obtained. The final step in the waiting time before the process is absorbed at 1 is observed to have an intriguing behaviour. The distribution of this final waiting time converges to the known distribution of the corresponding waiting time in the case of a single population as the migration rate tends to zero. The mean, however, does not converge. The waiting time until a sample has two common ancestors is modeled as a function of the migration rate. Finally bounds for the expected waiting time for the two colonies to have j > l ancestors are derived.


Lipoxygenase metabolites of arachidonic acid in neuronal transmembrane signalling.
Studies of invertebrate and vertebrate nervous tissue have demonstrated that free arachidonic acid and its lipoxygenase metabolites are produced in a receptor-dependent fashion. The intracellular actions of these compounds include the regulation of activity of membrane ion channels and protein kinases. In this article Daniele Piomelli and Paul Greengard review the evidence that these lipophilic molecules constitute a novel class of intracellular second messenger, possibly involved in the modulation of neurotransmitter release.


Does hepatitis C virus play a role in "non-viral" chronic liver disease?
It has recently been suggested that the hepatitis C virus may play a significant role in chronic liver diseases, such as autoimmune chronic active hepatitis, which are usually attributed to non-viral causes. We tested for antibodies to hepatitis C virus (anti-HCV) in sera from 140 patients with well characterised "non-viral" chronic liver diseases as well as sera from 51 patients thought to have chronic non-A, non-B (NANB) hepatitis (acting as positive controls) and 25 patients with non-hepatic autoimmune disorders. As expected, 45 of 51 patients (88%) diagnosed as having chronic NANB hepatitis were anti-HCV seropositive. Among 26 patients with cryptogenic cirrhosis, 8 were anti-HCV seropositive; in 5 patients (22%) there was no apparent risk factor for parenteral transmission. In the remaining 114 patients with chronic liver disease, 10 patients (9%) were seropositive for anti-HCV. However, 5 of these patients had a significant risk factor for parenteral transmission of hepatitis C virus, leaving only 5 of 106 (4.7%) with unexplained positive anti-HCV test results. Among patients with high titres of circulating autoantibodies but no liver disease, no positive results occurred. It is concluded that hepatitis C virus infection may account for some cases of cryptogenic cirrhosis. Although anti-HCV occurs more commonly in patients with other "non-viral" chronic liver diseases than has been reported in the community (0.5%-1.2%), the low prevalence of the antibodies indicates that hepatitis C virus infection is unlikely to be important in the aetiology or pathogenesis of autoimmune chronic active hepatitis and other poorly understood chronic liver diseases.


Novel stilbene-based Fischer base analog of leuco-TAM - (2E,2'Z)-{2-(4-(E)-styrylphenyl)propane-1,3-diylidene}bis(1,3,3-trimethylindoline) - derivatives: synthesis and structural consideration by 1D NMR and 2D NMR spectroscopy.
We report the synthesis of a series of novel stilbene-based (St) Fischer base analogs of leuco-triarylmethane (LTAM) dyes by treating Fischer base with (E)-4-styrylbenzaldehyde derivatives. All St-LTAM molecules examined herein are characterized by 1D and 2D NMR. They were found to exhibit ZE configuration and isomerize to their diastereomers EE and ZZ in 2-3 h. They exhibit type I behavior of diastereomeric isomerization.


Role of Polysaccharides Mimetic Components in Targeted Cancer Treatment.
Organic or inorganic compounds are synthesized or formulated to demonstrate their therapeutic actions, like a natural polysaccharide in the body. Polysaccharides, the major type of natural polymers, are biologically active, non-toxic, hydrophilic, and biodegradable and exhibit various properties. This manuscript is focused on delivering anticancer drugs with the help of mimetic components of polysaccharides. The data presented in this manuscript were obtained from PubMed, Elsevier, Taylor & Francis and Bentham Science Journals. Most chemotherapeutics are toxic to the human body, have a narrow therapeutic index, sluggish pharmaceutical delivery mechanisms, and are poorly soluble in water. The use of mimetic components of polysaccharides leads to the enhancement of the solubility of drugs in the biological environment. The current review summarizes the use of mimetic components of polysaccharides along with anticancer agents, which are capable of inhibiting the growth of cancerous cells in the body and exhibiting lesser adverse effects in the biological system compared to other therapies.


Chronic Constipation.
Chronic constipation is a common disorder that affects approximately 20% of the population and significantly impacts an individual's quality of life. The diagnosis can be made using standard criteria, and in the absence of alarm signs or symptoms, a determination of the underlying etiology/etiologies should be undertaken. In many instances, these will be gleaned from the history and physical examination. Specialized diagnostic testing may be warranted after the failure of initial laxative trials. Many therapeutic classes of laxatives exist with recent analyses indicating that practicing physicians prefer to use over-the-counter therapies in lieu of more strongly evidence-based prescription pharmaceuticals.


Obtaining drug exposure histories during pregnancy.
One hundred fifty-three pregnant patients were included in this study to verify the amount of drugs ingested during their pregnancies. The results demonstrate that, unknown to the physician, pregnant patients take a variety of pharmacologic agents. The patient's medical record is considered grossly inadequate in documenting an accurate assessment of a patient's exposure to drugs taken during pregnancy. Drug histories were obtained utilizing the hospital pharmacist drug history interview and home diary. Validity testing of the pharmacist's drug history and of the medical record history was not performed. The patient's compliance for utilizing the home diary for drugs or chemicals taken during their pregnancy was 83% (127 patients). It was shown that the usual methods of drug history documentation will identify only 30% of the actual drug exposure to the fetus.


Application of the PERMA Model of Well-being in Undergraduate Students.
The PERMA model was introduced by Seligman in 2011 to increase and measure well-being. This model defines well-being in terms of Positive Emotion, Engagement, Relationships, Meaning, and Accomplishment (PERMA). Mental health concerns are common in undergraduate students and may prevent them from obtaining optimal well-being. The purpose of this study was to test whether all five PERMA elements of well-being could be constructed from items within the 2018 Purdue University Student Experience at a Research University (SERU) survey. Using confirmatory factor analysis, all five PERMA constructs were supported and demonstrated good model fit statistics. A second order PERMA well-being construct was built and demonstrated adequate model fit with RMSEA = 0.04. All five constructs were significant at p < .001. Accomplishment had the highest factor loading (0.76) and Meaning had the lowest factor loading (0.25). Results for this study support use of well-being theory in the context of undergraduate students and provides enhanced understanding of well-being characteristics in this population.


Leaching characteristics of calcium-based compounds in MSWI Residues: From the viewpoint of clogging risk.
Leachate collection system (LCS) clogging caused by calcium precipitation would be disadvantageous to landfill stability and operation. Meanwhile, calcium-based compounds are the main constituents in both municipal solid waste incineration bottom ash (MSWIBA) and stabilized air pollution control residues (SAPCR), which would increase the risk of LCS clogging once these calcium-rich residues were disposed in landfills. The leaching behaviors of calcium from the four compounds and municipal solid waste incineration (MSWI) residues were studied, and the influencing factors on leaching were discussed. The results showed that pH was the crucial factor in the calcium leaching process. CaCO3 and CaSiO3 began leaching when the leachate pH decreased to less than 7 and 10, respectively, while Ca3(PO4)2 leached at pH<12. CaSO4 could hardly dissolve in the experimental conditions. Moreover, the sequence of the leaching rate for the different calcium-based compounds is as follows: CaSiO3>Ca3(PO4)2>CaCO3. The calcium leaching from the MSWIBA and SAPCR separately started from pH<7 and pH<12, resulting from CaCO3 and Ca3(PO4)2 leaching respectively, which was proven by the X-ray diffraction results. Based on the leaching characteristics of the different calcium compounds and the mineral phase of calcium in the incineration residues, simulated computation of their clogging potential was conducted, providing the theoretical basis for the risk assessment pertaining to LCS clogging in landfills.


Human resistin stimulates the pro-inflammatory cytokines TNF-alpha and IL-12 in macrophages by NF-kappaB-dependent pathway.
Resistin, a recently discovered 92 amino acid protein involved in the development of insulin resistance, has been associated with obesity and type 2 diabetes. The elevated serum resistin in human diabetes is often associated with a pro-inflammatory milieu. However, the role of resistin in the development of inflammation is not well understood. Addition of recombinant human resistin protein (hResistin) to macrophages (both murine and human) resulted in enhanced secretion of pro-inflammatory cytokines, TNF-alpha and IL-12, similar to that obtained using 5 microg/ml lipopolysaccharide. Both oligomeric and dimeric forms of hResistin were able to activate these cytokines suggesting that the inflammatory action of resistin is independent of its conformation. Heat denatured hResistin abrogated cytokine induction while treatment of recombinant resistin with polymyxin B agarose beads had no effect thereby ruling out the role of endotoxin in the recombinant hResistin mediated cytokine induction. The pro-inflammatory nature of hResistin was further evident from the ability of this protein to induce the nuclear translocation of NF-kappaB transcription factor as seen from electrophoretic mobility shift assays. Induction of TNF-alpha in U937 cells by hResistin was markedly reduced in the presence of either dominant negative IkappaBalpha plasmid or PDTC, a pharmacological inhibitor of NF-kappaB. A protein involved in conferring insulin resistance is also a pro-inflammatory molecule that has important implications.


Optimization of CLARITY for Clearing Whole-Brain and Other Intact Organs
The development, refinement, and use of techniques that allow high-throughput imaging of whole brains with cellular resolution will help us understand the complex functions of the brain. Such techniques are crucial for the analysis of complete neuronal morphology-anatomical and functional-connectivity, and repeated molecular phenotyping. CLARITY is a recently introduced technique that produces structurally intact, yet optically transparent tissue, which may be labeled and imaged without sectioning. However, the utility of this technique depends on several procedural variables during the process in which the light-scattering lipids in a tissue are replaced by a transparent hydrogel matrix. Here, we systematically varied a number of factors (including temperature, hydrogel composition, and polymerization conditions) to provide an optimized, highly replicable CLARITY procedure for clearing mouse brains. We found that for these preparations optimal tissue clearing requires electrophoresis (and cannot be achieved with passive clearing alone) for 5 d with a combination of 37 and 55°C temperature. Although this protocol is optimized for brains, we also show that it can be used to clear and analyze a variety of organs. Brain or other tissue prepared using this protocol is suitable for high-throughput imaging with confocal or single-plane illumination microscopy.


Acid-induced disassembly of the Haemophilus ducreyi cytolethal distending toxin.
The cytolethal distending toxins (CDTs) produced by many Gram-negative pathogens are tripartite genotoxins with a single catalytic subunit (CdtB) and two cell-binding subunits (CdtA + CdtC). CDT moves by vesicle carriers from the cell surface to the endosomes and through the Golgi apparatus en route to the endoplasmic reticulum (ER). CdtA dissociates from the rest of the toxin before reaching the Golgi apparatus, and CdtB separates from CdtC in the ER. The free CdtB subunit, which is only active after holotoxin disassembly, then crosses the ER membrane and enters the nucleus where it generates DNA breaks. We hypothesized that the acidified lumen of the endosomes is responsible for separating CdtA from the CdtB/CdtC heterodimer. To test this prediction, possible acid-induced disruptions to the CDT holotoxin were monitored by size exclusion chromatography and surface plasmon resonance. We found that CDT could not efficiently assemble from its individual subunits at the early endosome pH of 6.3. Partial disassembly of the CDT holotoxin also occurred at pH 6.3, with complete separation of CdtA from an intact CdtB/CdtC heterodimer occurring at both pH 6.0 and the late endosome pH of 5.6. Acidification caused the precipitation of CdtA at pH 6.5 and below, but neither CdtB nor CdtC were affected by a pH as low as 5.2. Circular dichroism further showed that the individual CdtB subunit adopts a different secondary structure as compared to its structure in the holotoxin. We conclude the first stage of CDT disassembly occurs in the early endosomes, where an acid-induced alteration to CdtA releases it from the CdtB/CdtC heterodimer.


Does the Z gene variant of alpha-1-antitrypsin predispose to hepatic carcinoma?
In 10 of 56 patients with primary liver carcinoma the nontumorous hepatocytes contained diastase resistant, periodic acid-Schiff positive and alpha-1-antitrypsin positive (immunoperoxidase technique) globules. This is a frequency of 18 per cent among patients with liver carcinoma against 6 per cent in an unselected autopsy series. The tumors were of the hepatocellular or mixed type in nine of the 10 patients, and the frequency among such patients was 23 per cent. We consider that these globules indicate a carrier of the protease inhibitor allele of the Z gene variant (single or double).


Tivantinib induces G2/M arrest and apoptosis by disrupting tubulin polymerization in hepatocellular carcinoma.
Tivantinib has been described as a highly selective inhibitor of MET and is currently in a phase III clinical trial for the treatment of hepatocellular carcinoma (HCC). However, the mechanism of tivantinib anti-tumor effect has been questioned by recent studies. We show that tivantinib indiscriminately inhibited MET dependent and independent HCC cells proliferation. In contrast, other MET inhibitors, JNJ-38877605 and PHA-665752, just specifically inhibited the growth of MET dependent HCC cells. Tivantinib neither inhibit constitutive MET phosphorylation nor HGF-induced MET phosphorylation in HCC cells. In the microtubule polymerization analysis, tivantinib affected microtubule dynamics by a mechanism as a microtubule depolymerizer. Interesting, unlike other microtubule-targeting agents, paclitaxel and vincristine, tivantinib showed similar anti-proliferative activity in parental and multidrug-resistant cells. Further studies demonstrated that tivantinib induced a G2/M arrest and promoted apoptosis by both intrinsic and extrinsic pathway. The in vivo efficacy evaluation showed that tivantinib exhibited a good anti-tumor growth activity with anti-proliferative and pro-apoptotic effects. The potent anti-tumor activity of tivantinib in HCC was achieved by targeting microtubule. Tivantinib treatment for patients with HCC should not be selected based on MET status.


Healthcare services gap analysis: a supply capture and demand forecast modelling, Dubai 2018-2030.
Health systems aim to provide a range of services to meet the growing demand of Dubai's heathcare system aims to provide a range of services to meet the growing demand of its population health needs and to ensure that standards of easy access, quality, equity and responsiveness are maintained. Dubai Health Authority (DHA) uses health services planning tools to assess the health needs of its population and sets priorities and effective regulatory strategies to achieve equilibrium of supply and demand of healthcare services and ensure adequate healthcare services are available, in terms of both quality and quantity. This study aims to measure the gap between demand and supply in health care services in Dubai at the baseline and to forecast the gap size and type (according to medical specialty, key medical planning units and geographical area) till 2030. The specific consequential aim includes identification of appropriate strategic directions for regulation, licensing, policies, insurance. The supply of healthcare services, professionals and medical equipment is captured through a census of all healthcare facilities licensed for practice in the Emirate of Dubai. The demand is estimated using a need based approach, where demand for episodes of medical care are estimated by age and gender and aligned to the internationally defined diagnosis related groups (IR-DGRs). The estimated episodes are then forecasted into the future, until 2030, using three scenarios of population growth (high, medium and low) for the emirate of Dubai. The captured supply and forecasted demand has been categorized into eight key health-planning units (KPUs) to allow for understanding of the population healthcare service needs by main service categories. Using a software for health services planning, a gap analysis between supply and demand is conducted till year 2030. The results revealed a current and expected undersupply and oversupply for some healthcare services by medical specialty and geographical area of the Emirate. By 2030, the largest gaps exists in acute beds, which would require 1,590 additional beds, for acute-same day beds, an additional 1575 beds, for outpatient consultation rooms, an additional 2,160 consultation rooms, for emergency department, an additional 107 emergency bays, and for long-term care and rehabilitation beds, an additional 675 beds. The top specialty needs for these categories include cardiology, orthopedics, rheumatology, psychiatry, pediatric medicine & surgery, gastroenterology, hematology & oncology, renal medicine, primary care, respiratory medicine, endocrinology, rehabilitation and long-term care. There is an existing and growing requirement to support the healthcare services capacity needs for the top service lines and geographical areas with the largest gaps. Future licensing is required to ensure that new facilities are geographically distributed in a balanced way, and requests for licensing that create or augment oversupply should be avoided.


Overexpression of Pygo2 Increases Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Cardiomyocyte-like Cells.
Our previous studies have shown that Pygo (Pygopus) in Drosophila plays a critical role in adult heart function that is likely conserved in mammals. However, its role in the differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into cardiomyocytes remains unknown. To investigate the role of pygo2 in the differentiation of hUC-MSCs into cardiomyocytes. Third passage hUC-MSCs were divided into two groups: a p+ group infected with the GV492-pygo2 virus and a p- group infected with the GV492 virus. After infection and 3 or 21 days of incubation, Quantitative real-time PCR (qRT-PCR) was performed to detect pluripotency markers, including OCT-4 and SOX2. Nkx2.5, Gata-4 and cTnT were detected by immunofluorescence at 7, 14 and 21 days post-infection, respectively. Expression of cardiac-related genes-including Nkx2.5, Gata-4, TNNT2, MEF2c, ISL-1, FOXH1, KDR, αMHC and α-Actin-were analyzed by qRT-PCR following transfection with the virus at one, two and three weeks. After three days of incubation, there were no significant changes in the expression of the pluripotency stem cell markers OCT-4 and SOX2 in the p+ group hUC-MSCs relative to controls (OCT-4: 1.03 ± 0.096 VS 1, P > 0.05, SOX2: 1.071 ± 0.189 VS 1, P > 0.05); however, after 21 days, significant decreases were observed (OCT-4: 0.164 ± 0.098 VS 1, P < 0.01, SOX2: 0.209 ± 0.109 VS 1, P < 0.001). Seven days following incubation, expression of mesoderm specialisation markers, such as Nkx2.5, Gata-4, MEF2c and KDR, were increased; at 14 days following incubation, expression of cardiac genes, such as Nkx2.5, Gata-4, TNNT2, MEF2c, ISL-1, FOXH1, KDR, αMHC and α-Actin, were significantly upregulated in the p+ group relative to the p- group (P < 0.05). Taken together, these findings suggest that overexpression of pygo2 results in more hUCMSCs gradually differentiating into cardiomyocyte-like cells. We are the first to show that overexpression of pygo2 significantly enhances the expression of cardiac-genic genes, including Nkx2.5 and Gata-4, and promotes the differentiation of hUC-MSCs into cardiomyocyte-like cells.


Resistance training in hemodialysis patients: a review.
Few studies have examined the role of resistance training (RT) in chronic kidney patients on hemodialysis (HD). This study reviews the literature about resistance exercise for patients on HD and describes protocols and clinical outcomes. A search of the MEDLINE database found 21 eligible publications, of which 14 studies applied only RT and 7 combined RT and aerobic training. Regarding the period of exercise training, 14 studies applied intradialytic exercise. The main outcome reported was muscle strength, which was assessed through knee strength and handgrip measures. All studies reviewed presented at least one significant result in relation to biochemical parameters, physical capacity, strength, body composition, or quality of life. This review demonstrates that RT improves muscle strength, some biochemical parameters, and quality of life of the HD patients.


Effect of exercise training on skeletal muscle fibre characteristics in men with chronic heart failure. Correlation between skeletal muscle alterations, cytokines and exercise capacity.
In patients with congestive heart failure (CHF) there is a shift from aerobic type I muscle fibres to less aerobic type II fibres. Exercise training has been shown to have beneficial effects on exercise performance, peripheral pathology and the neurohumoral profile in stable patients with CHF. This study evaluated the effect of a 3 month exercise training program on skeletal muscle characteristics and the correlation of these to cytokines and exercise capacity in CHF patients. Skeletal muscle biopsies for enzyme-histochemical analysis were performed in 15 CHF patients in New York Heart Association classes II-III, with a mean ejection fraction of 33+/-5% before and after a 12 week training period. The patients were trained for 30 min, five times a week at 80% of the peak heart rate achieved at baseline ergometer cycle test. Fifteen healthy men were used as controls. Plasma samples were examined by enzyme immunoassays for levels of pro-inflammatory cytokines. (a) At baseline we found muscle atrophy in five of the patients. The percent area of type I fibres (40.7+/-12.0 vs. 56.4+/-11.0%, P<0.05) and the thickness of type IIA (56.10+/-7.8 vs. 71.6+/-11.9 microm, P<0.001) and B-fibres (49.0+/-8.9 vs. 63.9+/-10.6 microm, P<0.001) were reduced, whereas the percent area of type IIA fibres (52.1+/-13.3 vs. 36.4+/-9.9%, P<0.05) was increased in heart failure patients compared to healthy controls. There was a modest correlation between fibre thickness and the level of interleukin 6 (r=-0.657, P=0.008). (b) After exercise training there was a reduction in muscle area examined by light-microscopy, measured as a percentage of field (-2.7, P=0.003) with an concomitant increase in interstitium. This reduction correlated to the increase in the 6-min walk test (r=-0.558, P=0.031). The thickness of type IIB fibres increased (+5.6 microm, P=0.068) and the area of type I fibres decreased (-6.1%, P=0.062). Patients with CHF have a relatively increased area of type IIA fibres and a relatively decreased area of type I fibres compared to healthy individuals. The thickness of type IIA and type IIB fibres is decreased compared to normal individuals. A modest negative correlation between the level of interleukin 6 and fibre thickness at baseline, suggests that inflammatory cytokines may be involved in the pathogenesis of the CHF related myopathy. A significant correlation between the reduction of muscle area, with increased interstitum, and the increase in the 6-min walk test may indicate that the improvement is due to increased capillary density permitting better flow reserve to exercising muscles.


Role of interleukin 2 on enhancement of concanavalin A-induced human peripheral blood lymphocyte proliferation by murine B cell mitogens.
Murine B cell mitogens such as bacterial lipopolysaccharide (LPS), butanol-extracted water soluble adjuvant (Bu-WSA), dextran sulfate (DS), synthetic muramyl dipeptide (MDP), and its analog MDP-Lys (L18) do not show any mitogenic ability in vitro on human peripheral blood lymphocytes or mixed cell populations of purified T and B cells obtained from the lymphocytes in an ordinary culture system. However, these mitogens are capable of enhancing the mitogenic effect of concanavalin A (Con A) in the cultures. In the presence of one of these mitogens, the activity of interleukin 2 (IL 2), but not interleukin 1, in the supernatants obtained from cultures containing Con A-stimulated T cell and B cell populations was higher than that of control cultures. The role of the newly produced IL 2 in the synergistic effect of the mitogens in human lymphocyte cell cultures was discussed.


Chronic Elevation of Tumor Necrosis Factor-α Mediates the Impairment of Leptomeningeal Arteriogenesis in db/db Mice.
Leptomeningeal collateral growth is a key factor that defines the severity of ischemic stroke. Patients with stroke generally have vascular risk factors, such as diabetes mellitus; however, consensus is lacking on how diabetes mellitus affects leptomeningeal arteriogenesis. We investigate the influence of diabetes mellitus on the leptomeningeal arteriogenesis. We measured the vessel diameter of the leptomeningeal anastomoses 14 days after the common carotid artery occlusion in db/db, db/+, and streptozotocin-induced hyperglycemic mice. In another set of these mice, we measured the infarct volume attributed to subsequent middle cerebral artery occlusion 14 days after the common carotid artery occlusion. Mac-2-positive cells on the dorsal brain surface and the mRNA expression of several macrophage-related factors in the cerebral cortex were examined. Finally, we tested whether the leptomeningeal arteriogenesis could be restored by pharmaceutical intervention in the db/db mice. Cerebral hypoperfusion led to significant ipsilateral leptomeningeal collateral growth in db/+ mice and streptozotocin-induced hyperglycemic mice. The collateral growth contributed to reduced infarct volume. In contrast, leptomeningeal arteriogenesis was impaired in the db/db mice. The number of Mac-2-positive cells was increased and tumor necrosis factor-α mRNA expression was induced after common carotid artery occlusion in the db/+ mice. However, these responses were not observed in the db/db mice. Administration of the tumor necrosis factor-α inhibitor etanercept before common carotid artery occlusion restored the hypoperfusion-induced leptomeningeal collateral growth in db/db mice. These results indicate that leptomeningeal arteriogenesis is impaired in db/db mice and that suppression of the tumor necrosis factor-α response to hypoperfusion is the major contributing factor.