Vaginal fluid urea and creatinine, an untapped diagnostic tool for premature rupture of membranes.
The diagnosis of premature rupture of membranes (PROM) can be difficult in equivocal cases. This study was designed to test the validity of vaginal fluid urea and creatinine in the diagnosis of PROM against the gold standard, the Amnisure ROM test™ METHODS: The study was a prospective observational study. All consenting eligible pregnant women between gestational ages of 28 weeks to 41 weeks + 6 days were recruited from the obstetrics emergency and antenatal clinic of the Federal medical centre, Keffi. Patients with history of drainage of liquor were recruited as the case group, and controls who match for age, parity and gestational age were recruited from the antenatal clinic to constitute the control group. Vaginal fluid aspirated was assessed in the laboratory for urea and creatinine levels and an Amnisure ROM test™ done. The sensitivity, specificity, negative and positive predictive values of vaginal fluid urea and creatinine were assessed in the diagnosis of PROM. Vaginal fluid urea and creatinine had sensitivity, specificity, negative predictive value (NPV), positive predictive value (PPV), respectively, of 94%, 82%, 93.18% and 83.93%, and 98%, 90%, 97.82% and 90.74%. The cutoff values for vaginal fluid urea and creatinine were 1.25 mg/dl and 0.23 mg/dl, respectively. This study has found that vaginal fluid urea and creatinine are very effective tests in diagnosis of PROM. It is a cheaper and more readily available alternative to the Amnisure test. It is especially useful in our environment, especially in cases of equivocal PROM, as a cost-effective means to confirm the diagnosis. 72961653, retrospectively registered on the 2020-07-09.


First report of an isolated jejunal seminoma: presentation with melaena and iron deficiency anaemia.
We present the case history of a man with isolated seminoma in the jejunum and abnormal testes but no provable malignant testicular disease. Treatment with cisplatin-based chemotherapy led to complete resolution of the jejunal seminoma. The rarity of seminoma involving the small bowel is highlighted. A literature search did not reveal other similar cases of isolated seminoma affecting the jejunum. The possible origins of this tumour are discussed.


The limiting factors of oncolytic virus immunotherapy and the approaches to overcome them.
Oncolytic virus (OV) immunotherapy is characterized by viruses which specifically target cancer cells and cause their cytolysis. They provide a unique and promising new tool for the eradication of cancer as they interact with and affect the tumor microenvironment (TME), vasculature, and immune system. Advancements of genetic engineering have allowed for these viruses to be armed in such a way to have enhanced targeting, strong immunomodulation properties, and an ability to modify the TME. However, there are still major limitations in their use, mostly due to difficulties in delivering the viral particles to the tumors and in ensuring that the immunomodulatory properties are able to stimulate the host immune response to mount a complete response. Using novel delivery systems and using OVs as a complementary therapy in a combinatorial treatment have shown some significant successes. In this review, we discuss the major issues and difficulties in using OVs as anti-tumor agents and some of the strategies put in place so far to overcome these limitations. KEY POINTS: • Oncolytic viruses (OVs) infect cancer cells and cause their cytolysis. • The major limitations in using OVs as anti-tumor therapy were discussed. • The potential strategies to overcome these limitations were summarized.


Radiochemotherapy in the management of pancreatic cancer--part II: use in adjuvant and locally unresectable settings.
Inherently, treating pancreatic adenocarcinoma with chemotherapy is limited by the marginal sensitivity of this tumor to available drugs and drug combinations. For radiotherapy, a similar therapeutic insensitivity is adversely combined with a compromised ability to escalate biologic dose within relevant target volumes because of an intertwining and surrounding matrix of dose-limiting critical normal organs and tissues. The need for effective adjuvant therapy has been intensified by an increase in the safety, applicability, and availability of surgical resection even though such resections remain largely inadequate for producing long-term disease-free survival. The need for improved treatments for locally unresectable tumors remains self-evident based on limited prognosis and the frequency of this presentation. Studies in both domains have attempted to incorporate technical advances in radiotherapy planning and delivery (3-dimensional conformal, intensity modulation) as well as the systemic promise and radiosensitizing properties of gemcitabine-based approaches. The challenges of clinical research are well reflected by the controversies resulting from available studies and the difficulties in successfully designing and executing future trials.


The Integration of Proteome-Wide PTM Data with Protein Structural and Sequence Features Identifies Phosphorylations that Mediate 14-3-3 Interactions.
14-3-3s are abundant proteins that regulate essentially all aspects of cell biology, including cell cycle, motility, metabolism, and cell death. 14-3-3s work by docking to phosphorylated Ser/Thr residues on a large network of client proteins and modulating client protein function in a variety of ways. In recent years, aided by improvements in proteomics, the discovery of 14-3-3 client proteins has far outpaced our ability to understand the biological impact of individual 14-3-3 interactions. The rate-limiting step in this process is often the identification of the individual phospho-serines/threonines that mediate 14-3-3 binding, which are difficult to distinguish from other phospho-sites by sequence alone. Furthermore, trial-and-error molecular approaches to identify these phosphorylations are costly and can take months or years to identify even a single 14-3-3 docking site phosphorylation. To help overcome this challenge, we used machine learning to analyze predictive features of 14-3-3 binding sites. We found that accounting for intrinsic protein disorder and the unbiased mass spectrometry identification rate of a given phosphorylation significantly improves the identification of 14-3-3 docking site phosphorylations across the proteome. We incorporated these features, coupled with consensus sequence prediction, into a publicly available web app, called "14-3-3 site-finder". We demonstrate the strength of this approach through its ability to identify 14-3-3 binding sites that do not conform to the loose consensus sequence of 14-3-3 docking phosphorylations, which we validate with 14-3-3 client proteins, including TNK1, CHEK1, MAPK7, and others. In addition, by using this approach, we identify a phosphorylation on A-kinase anchor protein-13 (AKAP13) at Ser2467 that dominantly controls its interaction with 14-3-3.


Striatal hypertrophy and its cognitive effects in new-onset benign epilepsy with centrotemporal spikes.
Benign epilepsy with centrotemporal spikes (BECTS), the most common childhood epilepsy syndrome, is a neurodevelopmental disorder with a genetic influence. Despite its signature electroencephalographic pattern and distinct focal motor seizure semiology, little is known about the underlying brain anatomic alteration and the corresponding cognitive consequences. Given the motor manifestations of seizures in BECTS, we hypothesize that anatomic networks in BECTS involve a distributed corticostriatal circuit. We investigated volumetric differences and shape deformities of caudate, putamen, pallidum, and thalamus in a group of children with new- and recent-onset BECTS (N = 3) compared to healthy controls (N = 54). We correlated specific subcortical volumes in BECTS that were significantly different from those in healthy controls with performances in executive function. Children with BECTS demonstrated significantly hypertrophied putamen, which was selective among the subcortical regions examined. Shape analysis showed dorsoventral elongation of the left caudate and bilateral putamen, with subnuclei expansion in ventral and dorsal striatum. Larger putamen volumes were linked to better cognitive performances on two complementary executive function tests. Children with BECTS showed aberrant volume and shape in subcortical regions that are critical for both motor processing and executive function. It is of importance to note that the hypertrophy appears to be cognitively adaptive, as enlargement was associated with improved cognitive performances. The anatomic abnormalities and their cognitive effects are evident in a group of children with new- and recent-onset epilepsy, suggesting that the structural brain anomalies occurred before the diagnosis of epilepsy.


Preparation and Study on Nickel Oxide Reduction of Polyacrylonitrile-Based Carbon Nanofibers by Thermal Treatment.
Carbon materials containing magnetic nanopowder have been attractive in technological applications such as electrochemical capacitors and electromagnetic wave shielding. In this study, polyacrylonitrile (PAN) fibers containing nickel nanoparticles were prepared using an electrospinning method and thermal stabilization. The reduction of nickel oxide was investigated under a nitrogen atmosphere within a temperature range of 600 to 1,000 °C. Carbon nanofibers containing nickel nanoparticles were characterized by FE-SEM, EDS, XRD, TGA, and VSM. It was found that nickel nanoparticles were formed by a NiO reduction in PAN as a function of the thermal treatment. These results led to an increase in the coercivity of nanofibers and a decrease in the remanence magnetization.


Ultraviolet Exposure Scenarios: Balancing Risks of Erythema and Benefits of Cutaneous Vitamin D Synthesis.
Exposure to sunlight is a major source of vitamin D for most people. Yet public health advice has focused overwhelmingly on avoiding exposure of unprotected skin because of the risks of erythema and skin cancer. Given that there are also health risks associated with low vitamin D status, we explore the possibilities of achieving a range of targets associated with vitamin D and the accompanying erythema risk. We have calculated the exposure required to gain a number of proposed oral-equivalent doses of vitamin D, as functions of latitude, season, skin type and skin area exposed, together with the associated risk of erythema, expressed in minimum erythema doses. The model results show that a recommended daily intake of 400 IU is readily achievable through casual sun exposure in the midday lunch hour, with no risk of erythema, for all latitudes some of the year, and for all the year at some (low) latitudes. We also show that such daily, sub-erythemal doses at lunchtime during the summer months is sufficient to avoid winter-time vitamin D deficiency for the UK all-weather climate, provided that lower arms and legs are exposed in the warmer months. At the higher proposed vitamin D dose of 1000 IU, lunchtime sun exposure is still a viable route to the vitamin but requires the commitment to expose greater areas of skin and is effective for a shorter period of the year. The highest vitamin D requirement considered was 4000 IU per day. For much of the globe and much of the year, this is not achievable in a lunchtime hour and where it is possible large areas of skin must be exposed to prevent erythema. When the only variable considered was skin type, latitudinal and seasonal limits on adequate vitamin D production were more restrictive for skin type 5 than skin type 2.


Intertidal macrofauna and environmental stress at a riverine-marine boundary.
A field experiment was carried out to test the effect of pore water salinity on the macrobenthic assemblages in an estuarine region of the Tagus estuary (Portugal) subjected to wide fluctuations in salinity. The conditions at the experimental site ranged from freshwater (minimum salinity 0.2) to mesohaline (maximum salinity 15.3). The experimental site was affected by an unexpected deposition of fluid mud during summer. Redundancy Analysis discriminated the experimental treatments along the first canonical ordination axis. The analysis also revealed an experimental gradient of increasing environmental stress, in which the minimal presence of organisms corresponded to treatments representing a high level of environmental stress. Sediment dynamics and saline fluctuations were the major factors that, together, determined the low macrofaunal abundance and species diversity at the experimental site. The most abundant macrofaunal species in this harsh environment were the polychaetes Hediste diversicolor and Streblospio shrubsolii.


Tubulin as a binding partner of the heag2 voltage-gated potassium channel.
The aim of this work was to investigate interactions of the human ether-a-go-go channel heag2 with human brain proteins. For this, we used heag2-GST fusion proteins in pull-down assays with brain proteins and mass spectrometry, as well as coimmunoprecipitation. We identified tubulin and heat shock 70 proteins as binding to intracellular C-terminal regions of the channel. To study functional effects, heag2 channels were expressed in Xenopus laevis oocytes for two-electrode voltage clamping. Coexpression of alpha-tubulin or the application of colchicine significantly prolonged channel activation times. Application at different times of colchicine gave similar results. The data suggest that colchicine application and tubulin expression do not affect heag2 trafficking and that tubulin may associate with the channel to cause functional effects. Coexpression of heat shock 70 proteins had no functional effect on the channel. The role of tubulin in the cell cytoskeleton suggests a link for the heag2 channel in tubulin-dependent physiological functions, such as cellular proliferation.


Changes in the proportions of plasma insulin, proinsulin and a higher-molecular-weight insulin during pre- and post-operative glucose-infusion tests.
1. Glucose-infusion tests were performed on patients admitted for elective upper abdominal surgery 1 day before and 1 day after operation. In addition to insulin and proinsulin, a third immunoreactive insulin species of mol. wt. 20 000--30 000 was detected in plasma from two patients. The heterogeneity of plasma immunoreactive insulin (IRI) and the need to consider the effects of all forms, including proinsulin and the high-molecular-weight species, is emphasized. 2. During preoperative glucose infusions there was an increase in the percentage of the total plasma IRI present as high-molecular-weight forms (i.e. proinsulin plus the species of mol. wt. 20 000--30 000) from 3.9% to 10.8%. On the first postoperative morning all patients showed an increase in the amounts of the heavier IRI types, which accounted for 13.9% of the total plasma IRI. 3. The changes in insulin and proinsulin are consistent with the release from the pancreas of an insulin/proinsulin mixture of constant proportions, and the longer circulating half-life of proinsulin. 4. Increases in the amounts of high-molecular-weight IRI species after surgery may have a partial role in the development of insulin resistance but are probably not a major determinant of the insulin-resistant state.


Hepatic artery infusion and chemoembolization in the management of liver metastases.
Hepatic metastases rather than the primary neoplasm usually dictate the course of the disease and patient's survival. For unresectable disease, intraarterial infusion of chemotherapy, embolization, and chemoembolization are viable alternatives. Intraarterial therapy for hepatic metastases is based on the dual blood supply of the normal liver (portal vein, 75%, and hepatic artery, 25%) and that of the tumors (hepatic artery, 90%). Intraarterial infusion delivers a higher concentration of chemotherapy, whereas chemoembolization adds ischemia and increased contact time with the tumor. Selective vascular occlusion for infusion, redistribution of the blood supply and pulsatile flow enhance the delivery of therapeutic agents to the liver.


The Relationship Between Testosterone-Replacement Therapy and Lower Urinary Tract Symptoms: A Systematic Review.
To systematically review prospective trials evaluating the clinical effects of testosterone-replacement therapy on lower urinary tract symptoms and prostate volume. We performed a literature review through PubMed, Embase, and Cochrane Library from 1994 to 2015 for prospective trials of hypogonadal men with benign prostatic hyperplasia or lower urinary tract symptoms treated with testosterone-replacement therapy. We evaluated the abstracts for outcomes related to International Prostate Symptom Score, prostate volume, and urodynamic parameters. An original cohort of 3079 abstracts was reviewed. Thirty-five trials were selected for inclusion. The majority of trials reviewed found no significant prostate growth due to testosterone-replacement therapy. Studies of men with baseline mild lower urinary tract symptoms demonstrated either no change or an improvement in symptoms after treatment. There was a lack of relevant urodynamic studies. Trials of men with the metabolic syndrome demonstrated uniform improvement in lower urinary tract symptoms. Forty-six percent of all the trials identified included exclusion criteria for baseline severe-range lower urinary tract symptoms or other signs of obstructive lower urinary tract symptoms. The current literature demonstrates scant support for a causative relationship between testosterone-replacement therapy, de novo or worsening lower urinary tract symptoms, and prostate volume. Furthermore, our review found an absence of high quality evidence that would support guideline recommendations that testosterone-replacement therapy is relatively contraindicated in men with severe-range lower urinary tract symptoms. Future clinical trials with more inclusive voiding criteria are needed.


Overexpression of a human prostate-specific glandular kallikrein, hK2, in E. coli and generation of antibodies.
The genomic and the cDNA clones of human glandular kallikrein (hK2), a member of the kallikrein family, have been isolated; however, the hK2 protein has not yet been identified and characterized. The deduced sequence of hK2 is highly homologous to prostate specific antigen (PSA), a widely accepted prognostic indicator of prostate carcinoma. Also, hK2 mRNA, like PSA mRNA, is exclusively expressed in prostatic epithelia. These two properties make hK2 a potentially useful marker for studying prostate cancer. In this paper, we describe for the first time the overexpression of the entire hK2 protein (pre-pro hK2:pphK2) in the E. coli system. Our system yields high levels of authentic pphK2 (as determined by partial amino acid sequence analysis) comprising about 40% of total cellular protein. pphK2 was purified to near homogeneity by preparative SDS/PAGE and used to generate anti-pphK2 antibodies in rabbits. The antibodies recognize the recombinant hK2 protein and a major band of approximately 34 kDa in seminal fluid.


Paraneoplastic cerebellar degeneration associated with anti-protein kinase Cgamma antibodies in a Chinese patient.
Anti-protein kinase Cgamma (anti-PKCγ) antibodies are rare onconeural antibodies associated with paraneoplastic cerebellar degeneration (PCD). To date, only two patients with PCD and anti-PKCγ antibodies have been reported. Here, we report a Chinese patient with PCD and anti-PKCγ antibodies. Screening for tumor revealed lymphoepithelial carcinoma in tonsil. The patient's symptoms improved gradually after radiotherapy for the lymphoepithelial carcinoma and intravenous immunoglobulin immunotherapy.


Modification methylase M.Sau3239I from Streptomyces aureofaciens 3239.
By chromatography on phosphocellulose and Heparin-Sepharose the modification methylase M.Sau3239I was detected and partly purified from cells of Streptomyces aureofaciens 3239. Methylation by this enzyme protects DNA from cleavage by the restriction endonuclease R.Sau3239I. The enzyme catalyzes methylation of adenine to N-6-methyladenine in the 5'-CTCGmAG-3' recognition sequence.


Early psychosis in the inner city: a survey to inform service planning.
In recent years, there has been a growing impetus to offer services for people with early psychosis. This paper describes a survey undertaken as the basis for the development of an early psychosis service. Rates of psychosis are known to differ between cities and rural areas and between ethnic groups. Identifying service needs is enhanced if the social and demographic characteristics, as well as the incidence, of people with psychosis in a given catchment area are known. The present study identified all presentations of people aged 16 or over with first and second episodes of psychosis in a 6-month period within a specified South London catchment area.A follow-up was carried out at 1 year. This survey confirmed existing knowledge on the social and service needs of this population, in finding high rates of unemployment and homelessness, poor engagement with services and frequent involuntary pathways to mental healthcare. Subgroups were also identified with specific needs, which are less often cited in the literature, such as groups of asylum seekers and people with young dependants. The results of the survey indicated that existing services were not adequately meeting the needs of people with early psychosis. They have been used to inform the development of an innovative service for people with early psychosis. The plans for this service are described.


HIV-1 reverse transcriptase targeted for proteasomal degradation as a prototype vaccine against drug-resistant HIV-1.
Acquisition of drug-resistance conferring mutations leads to an enhanced degradation of HIV-1 reverse transcriptase (RT) affecting its immunogenicity. The mechanism of this degradation is not known. We investigated the input of proteasome in this degradation, and explored a possibility to enhance the proteasomal degradation of RTs to potentiate the immunogenic peformance of RT genes. To this end, a C-terminal fusion was made of RT with ornithine decarboxylase (ODC) that is rapidly degraded by proteasome in an ubiquitine-independent fashion. Eukaryotic cells were transiently transfected with the genes for wild-type (wt) RT, multi-drug-resistant (MDR) RT, and their chimeras with ODC. RT expression in the presence or absence of the proteasome inhibitors MG132 and epoxomicin was quantified by Western blotting. Treatment with MG132 led to a two-fold increase in the level of wtRT, and a four-fold increase in the level of MDR-RT accumulation. Treatment with epoxomicin had virtually no effect on the accumulation of wtRT, while stabilizing MDR-RT two-fold. Since epoxomicin is a more specific proteasome inhibitor, it indicated that degradation of wtRT may not be solely proteasomal. Fusion to ODC considerably decreased the intracellular levels of both RT-ODC and MDR-RT-ODC as compared to parental proteins. MG132 treatment increased the intracellular RT-ODC content 20-fold (up the level of the MG132-treated wtRT; 60-80 fg/cell), and epoxomicin treatment, 10-fold as compared to non-treated samples. Thus, attachment of ODC moiety has modified the metabolic pathway of RT targeting it to proteasomal degradation. We are currently testing if this is translated into an enhanced MHC class I performance of wild-type and drug-resistant RTs in gene immunization.


A novel secreted protease from Pseudomonas aeruginosa activates NF-kappaB through protease-activated receptors.
The Pseudomonas aeruginosa-derived alkaline protease (AprA), elastase A (LasA), elastase B (LasB) and protease IV are considered to play an important role in pathogenesis of this organism. Although the sequence analysis of P. aeruginosa genome predicts the presence of several genes encoding other potential proteases in the genome, little has been known about the proteases involving in pathogenesis. Recently, Porphyromonas gingivalis gingipains and Serratia marcescens serralysin have been shown to activate protease-activated receptor 2 (PAR-2), thereby modulating host inflammatory and immune responses. Accordingly, we hypothesized that unknown protease(s) from P. aeruginosa would also modulate such responses through PARs. In this study, we found that P. aeruginosa produces a novel large exoprotease (LepA) distinct from known proteases such as AprA, LasA, LasB and protease IV. Sequence analysis of LepA showed a molecular feature of the proteins transported by the two-partner secretion pathway. Our results indicated that LepA activates NF-kappaB-driven promoter through human PAR-1, -2 or -4 and cleaves the peptides corresponding to the tethered ligand region of human PAR-1, -2 and -4 at a specific site with exposure of their tethered ligands. Considered together, these results suggest that LepA would require PARs to modulate various host responses against bacterial infection.


Gentamicin as a bactericidal antibiotic in tissue culture.
The effects of gentamicin on cellular physiology were studied in a total of 9 mammalian cell lines, using the following parameters: cell morphology and viability (cytotoxicity), proliferation, culture medium acidification, lactic acid production, lactate dehydrogenase release, virus susceptibility, and effects on karyotype. With regard to cytotoxicity no gross differences could be found in the sensitivity of the diploid and aneuploid cells investigated, as judged by morphological criteria. However, cells exposed to the antibiotic in the lag-log phase of growth showed damage at lower concentrations (1000 mug/ml) than cells treated in the stationary phase (2000 mug/ml). As regards the influence of gentamicin on cell growth and metabolsim, dose-response relationship were found proving that the antibiotic causes a depression of proliferation, a striking increase in lactate production, an elevated LDH release, and changes in pH behaviour. All these parameters were unaffected by concentrations up to 125 mug/ml. No gross changes in chromosome morphology and number could be detected in huploid cell line after 10 passages with 50 mug/ml gentamicin in lieu of the usual penicillin plus streptomycin combination. The minimal bactericidal concentrations (MBC) were determined in cell-free media and in tissue cultures against 4 species of bacteria. The MBC of gentamicin was generally lower as compared with the penicillin plus streptomycin combination. In some instances MBC was higher in the presence than in the absence of ti-sue culture cells. Comparison of the bactericidal efficiency against 31 strains of 7 species of bacteria of gentamicin (50 mug/ml) and penicillin plus streptomycin (100 units plus 100 mug/ml) in cell cultures proved that gentamicin is superior for control of bacterial growth in tissue culture.


Early intervention to promote medical student interest in surgery and the surgical subspecialties.
Concerns about projected workforce shortages are growing, and attrition rates among surgical residents remain high. Early exposure of medical students to the surgical profession may promote interest in surgery and allow students more time to make informed career decisions. The objective of this study was to evaluate the impact of a simple, easily reproducible intervention aimed at increasing first- and second-year medical student interest in surgery. Surgery Saturday (SS) is a student-organized half-day intervention of four faculty-led workshops that introduce suturing, knot tying, open instrument identification, operating room etiquette, and basic laparoscopic skills. Medical students who attended SS were administered pre-/post-surveys that gauged change in surgical interest levels and provided a self-assessment (1-5 Likert-type items) of knowledge and skills acquisition. First- and second-year medical students. Change in interest in the surgical field as well as perceived knowledge and skills acquisition. Thirty-three first- and second-year medical students attended SS and completed pre-/post-surveys. Before SS, 14 (42%) students planned to pursue a surgical residency, 4 (12%) did not plan to pursue a surgical residency, and 15 (46%) were undecided. At the conclusion, 29 (88%) students indicated an increased interested in surgery, including 87% (13/15) who were initially undecided. Additionally, attendees reported a significantly (p < 0.05) higher comfort level in the following: suturing, knot tying, open instrument identification, operating room etiquette, and laparoscopic instrument identification and manipulation. SS is a low resource, high impact half-day intervention that can significantly promote early medical student interest in surgery. As it is easily replicable, adoption by other medical schools is encouraged.


Pseudodystonia: A new perspective on an old phenomenon.
Pseudodystonia represents a wide range of conditions that mimic dystonia, including disorders of the peripheral nervous system, spinal cord, brainstem, thalamus, cortex and non-neurological conditions such as musculoskeletal diseases. Here, we propose a definition of pseudodystonia and suggest a classification based on underlying pathophysiological mechanisms. We describe phenomenology of different forms of pseudodystonia and point to distinctions between dystonia and pseudodystonia as well as challenging issues that may arise in clinical practice. The term pseudodystonia can be used to describe abnormal postures, repetitive movements or both, in which results of clinical, imaging, laboratory or electrophysiological investigations provide definite explanation of symptoms which is not compatible with dystonia. Pseudodystonia can be classified into non-neurological disorders of the musculoskeletal system, disorders of sensory pathways, disorders of motor pathways and compensatory postures in other neurological diseases. Presence of associated neurological findings in the affected body part is the key towards diagnosis of pseudodystonia. Additional supporting features are the presence of fixed postures, the absence of sensory trick, acute mode of onset and severe pain. Worsening on eye closure, traditionally considered typical for pseudodystonia, is not always present and can also appear in dystonia. It is challenging to separate dystonia and pseudodystonia in patients with thalamic lesions or corticobasal syndrome, where abnormal postures coexist with sensory loss. Many cases of pseudodystonia are treatable. Therefore, it is essential to consider pseudodystonia in a differential diagnosis of abnormal postures until a detailed neurological examination rules it out.


Influence of aging on the quantity and quality of human cardiac stem cells.
Advanced age affects various tissue-specific stem cells and decreases their regenerative ability. We therefore examined whether aging affected the quantity and quality of cardiac stem cells using cells obtained from 26 patients of various ages (from 2 to 83 years old). We collected fresh right atria and cultured cardiosphere-derived cells (CDCs), which are a type of cardiac stem cell. Then we investigated growth rate, senescence, DNA damage, and the growth factor production of CDCs. All samples yielded a sufficient number of CDCs for experiments and the cellular growth rate was not obviously associated with age. The expression of senescence-associated b-galactosidase and the DNA damage marker, gH2AX, showed a slightly higher trend in CDCs from older patients (≥ 65 years). The expression of VEGF, HGF, IGF-1, SDF-1, and TGF-b varied among samples, and the expression of these beneficial factors did not decrease with age. An in vitro angiogenesis assay also showed that the angiogenic potency of CDCs was not impaired, even in those from older patients. Our data suggest that the impact of age on the quantity and quality of CDCs is quite limited. These findings have important clinical implications for autologous stem cell transplantation in elderly patients.


Carbonic anhydrases--an overview.
Carbonic anhydrases (CAs, EC 188.8.131.52) are widespread metalloenzymes all over the phylogenetic tree, with at least 4 distinct gene families encoding for them. At least 16 different alpha-CA isoforms were isolated in mammals, where these enzymes play crucial physiological roles. Representatives of the beta-delta-CA family are highly abundant in plants, diatoms, eubacteria and archaea. These enzymes are efficient catalysts for the reversible hydration of carbon dioxide to bicarbonate, but at least the alpha-CAs possess a high versatility, being able to catalyze different other hydrolytic processes The catalytic mechanism of the alpha-CAs is understood in detail: the active site consists of a Zn(II) ion co-ordinated by three histidine residues and a water molecule/hydroxide ion. The latter is the active species, acting as a potent nucleophile. For beta- and gamma-CAs, the zinc hydroxide mechanism is valid too, although at least some beta-class enzymes do not have water directly coordinated to the metal ion. CAs are inhibited by two classes of compounds: the metal complexing anions and the sulfonamides and their isosteres (sulfamates, sulfamides etc.) possessing the general formula RXSO(2)NH(2) (R = aryl; hetaryl; perhaloalkyl; X = nothing, O or NH). At least 25 clinically used drugs/agents in clinical development show applications as diuretics and antiglaucoma drugs, anticonvulsants, with some compounds being developed as anticancer agents/diagnostic tools for tumors, antiobesity agents, and antimicrobials/antifungals (inhibitors targeting CAs from pathogenic organisms such as Helicobacter pylori, Mycobacterium tuberculosis, Plasmodium falciparum, Candida albicans, etc). Several important physiological and physio-pathological functions are played by CA isozymes present in organisms all over the phylogenetic tree, related to respiration and transport of CO(2)/bicarbonate between metabolizing tissues and the lungs, pH and CO(2) homeostasis, electrolyte secretion in a variety of tissues/organs, biosynthetic reactions, such as the gluconeogenesis and ureagenesis among others (in animals), CO(2) fixation (in plants and algae), etc. The presence of these ubiquitous enzymes in so many tissues and in so different isoforms, represents an attractive goal for the design of inhibitors or activators with biomedical applications.


High prevalence of vitamin D deficiency in 2-17 year olds presenting with acute fractures in southern Australia.
To determine vitamin D deficiency risk and other lifestyle factors in children aged 2-17 years presenting with an acute fracture to Sunshine Hospital. A prospective observational study was undertaken using a convenience sample data collected from children aged 2-17 years of age presenting with an acute fracture. Recruitment was undertaken over a 3-month period from February to May 2014. Risk factors for vitamin D deficiency (skin pigmentation, hours spent outdoors, sunscreen use and obesity) were identified. Patients providing consent, had measurements of serum 25-hydroxyvitamin D (25-OHD). Vitamin D deficiency was defined as < 50 nmol/L. Of the 163 patients recruited into this study, 134 (82%) had one or more risk factor(s) for vitamin D deficiency. Of these, 109 (81%) consented to 25-OHD testing, with a median of 53 nmol/l (range 14-110 nmol/l) obtained. A total of 57 (52% at risk, 35% of total participants) were found to be vitamin D deficient. 45 (80%) had mild deficiency (30-50 nmol/l) and 11 (20%) had moderate deficiency (12.5-29 nmol/l). One third of all participants, and the majority participants who had one or more risk factor(s) for vitamin D deficiency, were vitamin D deficient. Based on our findings we recommend that vitamin D status be assessed in all children with risk factor of vitamin D deficiency living in urban environments at higher latitudes presenting with fractures. The effect of vitamin D status on fracture risk and fracture healing in children and teenagers is yet to be determined, as do the effects of vitamin D supplementation in vitamin D deficient paediatric patients presenting with acute fracture.


Situs inversus totalis: the association of Kartagener's syndrome with diffuse bronchiolitis and azoospermia.
Kartagener's syndrome (KS) is characterised by bronchiectasis, paranasal sinusitis and situs inversus totalis (SIT). Association of diffuse bronchiolitis (DB) with KS has been documented from Japan only. Fourteen patients with SIT were seen in one unit over 7 years. All patients underwent a similar work up which included high resolution computed-tomography (HRCT) of thorax and CT-paranasal sinuses. Semen analysis was done in 2/3 adult males. Eleven patients (6 males and 5 females) had KS while 3 adult females had SIT alone. HRCT-thorax detected bronchiectasis in 10/11 patients with KS. HRCT-thorax confirmed DB in 6/11 patients with KS. One adult male had total sperm count of 2.5 million/ml without sperm motility while the other had no sperms. This series documents the largest number of patients with SIT/KS from India and highlights for the first time, the association of DB with KS from India.


Brain-spleen axis in health and diseases: A review and future perspective.
The spleen, an important tissue for the immune system, acts as a filter for blood within the immune system. Accumulating evidence suggests that the spleen affects a number of brain functions in health and diseases via immune modulation. Systemic inflammation or chronic social defeat stress (CSDS) can cause splenomegaly in rodents. Interestingly, the new antidepressant arketamine could normalize splenomegaly and depression-like behaviors in CSDS-susceptible mice. A recent study strongly supports the direct connection pathway between the brain and spleen, whereby the spleen can regulate the humoral immune defense by the two brain regions, such as corticotropin-related neurons in the paraventricular nucleus (PVN) and the central nucleus of the amygdala (CeA). Furthermore, afferent and efferent vagus nerve signaling may contribute to brain and spleen communication. In this article, we review recent findings of the brain-spleen axis in health and diseases.


Can cerebral hypoperfusion after sympathetic storm be used to diagnose brain death? A retrospective survey in traumatic brain injury patients.
The aim of this study was to identify the precise time of occurrence of irreversible coma from brain stem dysfunction that precedes brain death. Sympathetic storm, which is a cardiovascular hyperdynamic state manifested by brain stem ischemia, is known to be related to brain stem failure in animal models. We studied sympathetic storm in the clinical setting and compared the clinical findings observed before and after sympathetic storm to identify the precise time of occurrence of irreversible apneic coma. We conducted a retrospective study of 15 comatose traumatic brain injury adult patients at the National Taiwan University Hospital's Neurosurgical Intensive Care Unit. Data on arterial blood pressure, heart rate, intracranial pressure, and clinical findings such as cerebral blood flow pattern, Glasgow Coma Scale, brain stem reflexes, utilizations of catecholamines, and occurrence of central diabetes insipidus throughout the course in the intensive care unit were collected retrospectively from medical records. Prolonged uncorrectable cerebral hypoperfusion was found after a characteristic irreversible apneic coma-associated sympathetic storm (IACASS) in all 15 patients. A mean cerebral perfusion pressure of 11.9 mm Hg +/- 10.3 mm Hg and 13 mm Hg +/- 3.5 mm Hg remained at 12 hours and 24 hours, respectively, after IACASS. Differences in clinical findings before and after IACASS that were statistically significant were cerebral circulation pattern (p = 0.0455), Glasgow Coma Scale (p = 0.0143), brain stem reflexes (p = 0.0143), utilization of catecholamines (p = 0.0254), and occurrence of central diabetes insipidus (p = 0.00468). Coma might have become irreversible immediately after IACASS because the prolonged duration of cerebral hypoperfusion could have caused irreversible cerebral tissue injury. Our study provides some preliminary findings suggesting that IACASS may be a predictor of impending brain death. A prospective study is the next step to understanding whether this phenomenon can be applied clinically to diagnose irreversible apneic coma.


Copper incorporation into recombinant CotA laccase from Bacillus subtilis: characterization of fully copper loaded enzymes.
The copper content of recombinant CotA laccase from Bacillus subtilis produced by Escherichia coli cells is shown to be strongly dependent on the presence of copper and oxygen in the culture media. In copper-supplemented media, a switch from aerobic to microaerobic conditions leads to the synthesis of a recombinant holoenzyme, while the maintenance of aerobic conditions results in the synthesis of a copper-depleted population of proteins. Strikingly, cells grown under microaerobic conditions accumulate up to 80-fold more copper than aerobically grown cells. In vitro copper incorporation into apoenzymes was monitored by optical and electron paramagnetic resonance (EPR) spectroscopy. This analysis reveals that copper incorporation into CotA laccase is a sequential process, with the type 1 copper center being the first to be reconstituted, followed by the type 2 and the type 3 copper centers. The copper reconstitution of holoCotA derivatives depleted in vitro with EDTA results in the complete recovery of the native conformation as monitored by spectroscopic, kinetic and thermal stability analysis. However, the reconstitution of copper to apo forms produced in cultures under aerobic and copper-deficient conditions resulted in incomplete recovery of biochemical properties of the holoenzyme. EPR and resonance Raman data indicate that, presumably, folding in the presence of copper is indispensable for the correct structure of the trinuclear copper-containing site.


Duck lymphocytes. II. Culture conditions for optimum transformation response to phytohaemagglutinin.
The lymphocyte transformation (LT) test was performed with duck blood lymphocytes in 96-well microtitre trays against nine concentrations (0.1-100 micrograms/ml) of phytohaemagglutinin (PHA). It was found that incubation at 41.6 degrees C (duck body temperature) gave better results than incubation at 37 degrees C, and that a 6 h pulse with [3H]thymidine gave better results than an 18 h pulse. Cell concentration was very critical: successful duck LT required rather high concentrations of cells (approximately 4 X 10(6)/ml, equivalent to approximately 8 X 10(5)/culture) while lower and higher concentrations gave inferior results. Duck serum supported LT optimally when used at a concentration of 10-15%, but a serum pool from adult (approximately 6-month-old) ducks gave superior results to sera from younger birds. Foetal calf serum supported duck LT strongly when used at 20%, but only poorly at 10%, while chicken serum supported duck LT when used at 5-20%. Ultroser G was an ineffective supplement, and the addition of 5% tryptose phosphate broth to other supplements had variable effects. Using optimum culture conditions, maximum LT responses were obtained after 2-3 days' culture, when it was possible to obtain stimulation indices of 500-5000. The addition of PHA to cultures could be delayed for up to 18 h without affecting LT, but thereafter the cells rapidly lost responsiveness.


Autolytic activity of human calpain 7 is enhanced by ESCRT-III-related protein IST1 through MIT-MIM interaction.
Calpain 7, a mammalian ortholog of yeast Cpl1/Rim13 and fungal PalB, is an atypical calpain that lacks a penta-EF-hand domain. Previously, we reported that a region containing a tandem repeat of microtubule-interacting and transport (MIT) domains in calpain 7 interacts with a subset of endosomal sorting complex required for transport (ESCRT)-III-related proteins, suggesting involvement of calpain 7 in the ESCRT system. Although yeast and fungal calpains are thought to be involved in alkaline adaptation via limited proteolysis of specific transcription factors, proteolytic activity of calpain 7 has not been demonstrated yet. In this study, we investigated the interaction between calpain 7 and a newly reported ESCRT-III family member, increased sodium tolerance-1 (IST1), which possesses two different types of MIT-interacting motifs (MIM1 and MIM2). We found that glutathione-S-transferase (GST)-fused tandem MIT domains of calpain 7 (calpain 7MIT) pulled down FLAG-tagged IST1 expressed in HEK293T cells. Coimmunoprecipitation assays with various deletion or point mutants of epitope-tagged calpain 7 and IST1 revealed that both repetitive MIT domains and MIMs are required for efficient interaction. Direct MIT-MIM binding was confirmed by a pulldown experiment with GST-fused IST1 MIM and purified recombinant calpain 7MIT. Furthermore, we found that the GST-MIM protein enhances the autolysis of purified Strep-tagged monomeric green fluorescent protein (mGFP)-fused calpain 7 (mGFP-calpain 7-Strep). The autolysis was almost completely abolished by 10 mmN-ethylmaleimide but only partially inhibited by 1 mm leupeptin or E-64. The putative catalytic Cys290-substituted mutant (mGFP-calpain 7(C290S)-Strep) showed no autolytic activity. These results demonstrate for the first time that human calpain 7 is proteolytically active, and imply that calpain 7 is activated in the ESCRT system.


Stroke associated with alpha blocker therapy for benign prostatic hypertrophy.
Benign enlargement of the prostate is a malady of older males, reaching an estimated prevalence of 90% in patients aged over 70 years. Many of these patients are treated with alpha blockers, which can lower blood pressure significantly. We report on a 64-year-old man who developed a right hemiparesis after taking one dose of doxazosin 4 mg for prostatic symptoms. A CT scan of the brain and carotid ultrasound studies were normal. He recovered most of his neurological function within a few days. Ambulatory blood pressure monitoring on doxazosin 2 mg revealed a striking sleep blood pressure reduction.


A novel classification of alar retraction based on nostril exposure in Asian rhinoplasty.
Alar retraction is considered a challenge in rhinoplasty. The classification of alar retraction remains poorly defined, especially in Asia. Patients with alar retraction are associated with excessive exposure to the nostrils in Asia. This study aimed to introduce a classification method of alar retraction based on nostril exposure. Medical records of patients who had undergone rhinoplasty with alar retraction based on nostril exposure between January 2015 and December 2020 were retrospectively reviewed. The corrections of alar retraction were categorized into three groups according to a classification method of alar retraction based on nostril exposure: mild alar retraction, moderate alar retraction, and severe alar retraction. The visual analog scale (VAS) and rhinoplasty outcomes evaluation (ROE) were used to evaluate the satisfaction rate. Within a median period of 13.3 months, 45 patients (51.14%) with mild alar retraction were corrected by alar contour graft. 23 patients (26.14%) with moderate alar retraction were treated with the alar contour graft(n=10), the lateral crural strut graft (n = 6), and the alar projection graft (n = 7). 20 patients (22.73%) with severe alar retraction were corrected by lateral crural strut graft combined with composite graft (n = 6), lateral crural strut graft (n = 10), and composite graft (n = 4). The severe alar retraction group had a higher satisfaction rate in ROE(P<0.05) and VAS (P<0.05) than moderate alar retraction and mild alar retraction at a follow-up of 12 months after surgery. The classification of alar retraction based on nostril exposure is more practical for rhinoplasty in Asia.


Fluorescence of equine platelet tropomyosin labeled with acrylodan.
Equine platelet tropomyosin was labeled with the sulfhydryl-specific fluorescent reagent 6-acryloyl-2-dimethylaminonaphthalene (acrylodan). The extent of labeling at 4 degrees C could be regulated between 0.5 and 1.3 acrylodans per tropomyosin chain by varying the reaction time from 1 to 4.5 h. Acrylodan-labeled platelet tropomyosin, AD-P-TM, was highly fluorescent, having an emission maximum near 518 nm on excitation at 365 nm. Steady-state measurements of polarization of the fluorescence of AD-P-TM in both low and high ionic strength solutions gave Perrin plots that exhibited sharp changes in slope near 50 degrees C, indicative of a sharp increase in mobility of the label at that temperature. This correlates with the melting temperature of the platelet tropomyosin coiled coil observed by circular dichroism [G. P. Côté, W. G. Lewis, M. D. Pato, and L. B. Smillie, (1978) FEBS Lett. 91, 237-241]. Perrin plots of carboxypeptidase A-treated platelet tropomyosin that was labeled with acrylodan after digestion resembled more closely those of acrylodan-labeled cardiac tropomyosin rather than those of AD-P-TM, suggesting that the observed emission arose from label at Cys-153 on each truncated platelet tropomyosin chain. In solutions containing 150 mM KCl and 5 mM MgCl2, addition of actin at up to a sixfold molar excess over AD-P-TM caused both the fluorescence emission intensities and fluorescence polarization values of samples to increase. In the presence of actin, the wavelength of maximal emission was shifted to shorter values by about 5 to 7 nm. These changes indicate that actin does bind to AD-P-TM and that the binding affects the environment of the label, both by making it more hydrophobic and by reducing the freedom of the label to tumble in solution.


On psychiatric symptoms of neuro-Behçet's syndrome.
Studies were made on the groups of psychiatric symptoms of Neuro-Behçet's syndrome in six cases considered to be brain stem encephalitis, including one case of autopsy. Psychiatric symptoms appeared from one to eight years after neurological symptoms first occurred. This group of psychiatric symptoms can be divided roughly into the early-stage, middle-stage, terminal-stage and episodic groups. The initias-stage symptom group concerns mainly a change in, and regression of, personality, consisting of being childish, shallow, indifferent, careless, rude, slow in action, euphoric, or depressive. The middle-stage symptom group is made up of indifference, forced smiling, forced crying, amnesia, recent memory impairment, and disorder of sleep rhythm. The terminal-stage symptom group comprises the "lock-in" syndrome, which if advanced, will become Apallisches Syndrome. The episodic symptom group is made up of pathologically abnormal experiences such as hallucinations and delusions and symptoms due to a decline in the level of consciousness such as confusion, delirium and twilight state. Dementia hitherto mentioned in Neuro-Behçet's syndrome should be classified as subcortical dementia.


Validation of the SACOV-19 score for identifying patients at risk of complicated or more severe COVID-19: a prospective study.
Identification of patients at risk of complicated or more severe COVID-19 is of pivotal importance, since these patients might require monitoring, antiviral treatment, and hospitalization. In this study, we prospectively evaluated the SACOV-19 score for its ability to predict complicated or more severe COVID-19. In this prospective multicenter study, we included 124 adult patients with acute COVID-19 in three German hospitals, who were diagnosed in an early, uncomplicated stage of COVID-19 within 72 h of inclusion. We determined the SACOV-19 score at baseline and performed a follow-up at 30 days. The SACOV-19 score's AUC was 0.816. At a cutoff of > 3, it predicted deterioration to complicated or more severe COVID-19 with a sensitivity of 94% and a specificity of 55%. It performed significantly better in predicting complicated COVID-19 than the random tree-based SACOV-19 predictive model, the CURB-65, 4C mortality, or qCSI scores. The SACOV-19 score is a feasible tool to aid decision making in acute COVID-19.


Cerebrospinal fluid amine metabolites after combined amitriptyline-triiodothyronine treatment of depressed women.
Levels of 5-hydroxyindoleacetic acid and homovanillic acid were measured in cerebrospinal fluid from 33 depressed women with no clinical response to amitriptyline, before and after combination treatment with triiodothyronine. Although the latter showed significant clinical improvement, changes in CSF amine metabolites did not differ significantly from a control group of 16 therapy-resistant depressed women receiving higher doses of amitriptyline. Possible explanations for the mechanism of action of triiodothyronine are discussed.


Outcome of Heart Transplant Candidates Based on a High Volume Heart Transplant Center Experience in Iran.
The waiting list for heart transplants is a valuable data registry that would offer very useful information on the characteristics of patients who have various outcomes while waiting in the list. The purpose of this study was to look at the prognosis of those waiting for heart transplants as well as the factors that increase mortality. Advanced heart failure patients' demographic, clinical, hemodynamic, and echocardiographic results, as well as their prognosis, were retrieved from the national registry for heart transplantation between 2011 and 2018. The study population was defined and compared in four groups: 1) Death while awaiting HTX, 2) Death after HTX, 3) Alive without a transplant, 4) Transplanted and alive. The data of 207 patients [75% male, mean (SD) age of 34(10) years] were analyzed. The most common etiology of heart failure was idiopathic dilated cardiomyopathy. A total of 86 patients (41%) were successfully transplanted, with a median (IQR) time between listing and transplantation of 84 (30¬219) days, 54 patients (26.1%) were dead and 32% were still alive. The multivariate analysis showed right atrial pressure, pulmonary capillary wedge pressure, cardiac index, and systolic blood pressure at the time of listing as independent predictors of death. The study on HTX waiting list is very useful for both allocation strategies and administrative planning for patients with advanced heart failure by development of accurate models and scoring systems using predictors of death in the waiting list.


Body mass index trajectories from childhood concerning emotional states in young adulthood: Tehran Lipid and Glucose Study.
This study aimed to identify body mass index (BMI) trajectories from childhood and their relationships with depression, anxiety, and stress symptoms in young adulthood. A total of 687 children aged 4-18 years were recruited from the Tehran Lipid and Glucose Study. Throughout 18 years of follow-up, BMI was measured every 3 years for a maximum of 6 data points. Participants completed the depression, anxiety, and stress scale in their young adulthood (aged 22-36). The group-based trajectory modelling was applied to identify BMI patterns. The logistic regression model was used to assess the association between BMI trajectories and depression, anxiety, and stress symptoms in adulthood. Two BMI trajectories were identified from childhood to young adulthood: healthy weight (HW = 69.6%) and persistent increasing overweight/obesity (PIO = 30.4%). After adjusting for potential confounders, compared with the HW group, men in the PIO group were more likely to experience higher stress levels (OR: 1.62, 95% CI: 0.99-2.63; p = 0.05). No significant association was observed between the PIO trajectory and depression and anxiety among both sexes and stress symptoms in females. Our results highlight that developing overweight and obesity from childhood may be related to increased stress in males.


Important role of blood rheology in atherosclerosis of patients with hemodialysis.
Concerning a role of blood rheology for atherosclerosis in patients with hemodialysis (HD), little data are available. It may be due to the fact that the method for evaluating rheologic properties of circulating blood has been limited. We examined blood rheology in 118 HD patients by using microchannel array flow analyzer that makes it possible to directly observe the flow of blood cell elements through the microchannel. Transit time (T(B)) of heparinized whole blood through slit pores (7 x 30 microm) was used as an index of rheology and related with various inflammatory biomarkers such as high-sensitive CRP (hsCRP), monocyte chemotactic protein-1, osteopontin, or fibrinogen (Fg). Moreover, as a surrogate marker of atherosclerosis, carotid intima-media thickness (IMT) and aortic stiffness evaluated by brachial-ankle pulse-wave velocity (baPWV) were studied. In HD patients, T(B) had strong positive correlations with hsCRP (r = 0.427; p < 0.00001), Fg (r = 0.452; p < 0.00001), and osteopontin (r = 0.227; p < 0.0134). Further, T(B) was significantly well correlated with IMT (r = 0.400; p < 0.0001) and PWV (r = 0.470; p < 0.0001). Multivariate regression analysis showed that baPWV, IMT, Fg, hematocrit, white blood cell count, and CRP were chosen as significant explanatory factors for T(B.) These results suggest that blood rheology may play an important role for atherosclerosis in patients with HD.


Case Study: Systematic Detection and Prioritization of Gene Fusions in Cancer by RNA-Seq: A DIY Toolkit.
RNA-seq provides an efficient and sensitive methodology to identify fusion transcripts in cancer tissues. Chimeric reads mapping across two different genes represent potential gene fusions. Various methodologies have been implemented in the detection of gene fusions by RNA-seq. Here we describe a general methodology used in processing and filtering of RNA-seq data, followed by filtering of multiple varieties of artifacts to nominate potentially relevant gene fusions. Functional relevance of gene fusions is assessed based on the predicted domain architecture of the putative fusion proteins.


New macroscopic classification and back-flow thrombectomy for advanced hepatocellular carcinoma with portal vein tumor thrombus invading the contralateral second portal branch.
Patients with hepatocellular carcinoma (HCC) and portal vein tumor thrombus (PVTT) invading the portal trunk (Vp4) are poor surgical candidates because of the technical difficulties involved. To overcome the limitations, we developed a technique of back-flow thrombectomy (BFT) based on the inherent portal hemodynamics and the macroscopic form of PVTT. Forty-six patients with multiple HCC and Vp4 PVTT underwent hepatectomy with tumor thrombectomy. We used the BFT to treat 24 patients, 18 of whom had PVTT in the contralateral second portal branch. The form of PVTT was classified macroscopically into the floating and expansive types. The rate of complete removal by BFT of PVTT in the contralateral second portal branch was 89%. The patency rates at the thrombectomy site in all 46 patients and in the 24 BFT patients, 3 months after hepatectomy were 93 and 90%, respectively. The median OS of all 46 patients was 15 months, with 1- and 3-year OS rates of 58.5 and 17.1%, respectively. The median OS of the 24 patients treated with BFT vs. the 22 not treated with BFT was 14 and 15 months, respectively. BFT can expand the therapeutic time window for patients with HCC and deep-seated PVTT and may improve their survival.


The content of psychoactive phenylpropyl and phenylpentenyl khatamines in Catha edulis Forsk. of different origin.
Khat, the leaves of Catha edulis Forsk. (Celastraceae), is widely used as a stimulant in East Africa and the Arab Peninsula. Samples from the most important markets of Ethiopia, Kenya, North Yemen and Madagascar were analysed with high-performance liquid chromatography (HPLC) concerning their khatamine content (phenylpropyl and phenylpentenylamines). The variability of khatamines depending on origin, type and quality is discussed. In many samples a good correlation between the amount of cathinone, the main CNS-active compound, and quality estimation (price) of dealers and consumers was found.


Diphtheria toxin fused to variant interleukin-3 provides enhanced binding to the interleukin-3 receptor and more potent leukemia cell cytotoxicity.
Chemoresistance is a common cause of treatment failure in patients with acute myeloid leukemia (AML). We generated a diphtheria toxin (DT) fusion protein composed of the catalytic and translocation domains of DT (DT388) fused to interleukin-3 (IL-3). IL-3 receptors (IL-3R) are overexpressed on blasts from many AML patients. DT388IL-3 showed cytotoxicity to leukemic blasts in vitro and in vivo and minimal damage to normal tissues in nonhuman primate models. However, only a fraction of patient leukemic samples were sensitive to the agent. To enhance the potency and specificity of the DT388IL-3 molecule, we constructed variants with altered residues in the IL-3 moiety. Two of these variants, DT388IL-3[K116W] and DT388IL-3[Delta125-133], were produced and partially purified from Escherichia coli with excellent yields. They showed enhanced binding to the human IL-3R and greater cytotoxicity to human leukemia cell lines relative to wild-type DT388IL-3. Interestingly, the results support a previously hypothesized model for interaction of the C-terminal residues of IL-3 with a hydrophobic patch on the alpha-subunit of IL-3R. Rational modification of the targeting domain based on structural analysis can produce a fusion toxin with increased ability to kill tumor cells. One or both of these variant fusion proteins merit further development for therapy of chemotherapy refractory AML.


Depressive symptoms and the implicit evaluation of alcohol: the moderating role of coping motives.
Depressive symptoms in college students have been associated with a number of indices of hazardous drinking. Investigators have utilized a variety of experimental paradigms to better understand the cognitive-motivational mechanisms that may underlie this association. Implicit cognition studies have provided increasing support for the view that coping motives may moderate the association between negative affect and the incentive value of alcohol. However, less is known about how symptoms of depression may be linked with implicit evaluative responses to alcohol. The current research sought to investigate the association between depressive symptoms and implicit evaluations of alcohol stimuli as measured by an evaluative priming task. Eighty-two current drinkers completed assessments of depressive symptoms, alcohol use, and drinking motives before engaging in a computerized alcohol evaluative priming task. Analyses showed that the association between depressive symptoms and the positive implicit evaluation of alcohol was moderated by drinking motives, such that the positive association between depression and alcohol evaluation was stronger for students who had higher coping motives. These findings add to our understanding of how depressive symptoms may influence responses to alcohol and provide further support for the view that individual differences in coping motives may be a critical moderator of the association between depressive symptoms and alcohol-related cognitive motivational processes.


[Variants of chromosome compactization and the estimation of the capacity of mammalian oocytes for further development].
The morphology of chromatin in spontaneously ovulated rat's oocytes was investigated. The degree of chromatin condensation was shown to vary widely: along with the typical cytological picture of metaphase II, both highly condensed (clamped) and decondensed ("not typical") forms of chromosomes were observed. A direct relation was discovered between the prevalence of the not typical morphological forms of oocyte chromosomes and the level of embryonal death. The shape of clamped and decondensed forms increased in the oocytes from old rats and in post-ovulatory aged oocytes.


[Drug-induced pneumonitis due to sirolimus: an interaction with atorvastatin?].
Sirolimus is an immunosupressant used in renal transplantation because of its lack of nephrotoxicity. We report four cases of pneumonitis due to sirolimus, possibly revealing an interaction with atorvastatin. Four patients (previously on long-term treatment with atorvastatin) presented with respiratory symptoms between 3 and 56 months after starting treatment with sirolimus following renal transplantation. Thoracic CT scans showed bilateral areas of peripheral alveolar consolidation. Bronchial lavage showed a lymphocytic alveolitis. Open-lung biopsy showed organizing pneumonia associated with diffuse alveolar damage and caseating granulomata. We attributed the pneumonitis to sirolimus on account of clinical and radiological resolution within 1 to 6 months of stopping treatment. We raise the possibility of an association between sirolimus and atorvastatin by competition for their hepatic degradation pathway via cytochrome P450 3A4. Sirolimus causes drug-induced pneumonitis that is predominantly an organizing pneumonia. Atorvastatin may encourage its development by competition with sirolimus in the liver.


Corrigendum: A Novel Phage PD-6A3, and Its Endolysin Ply6A3, With Extended Lytic Activity Against Acinetobacter baumannii.
[This corrects the article DOI: 10.3389/fmicb.2018.03302.].


Neurotransmitter vesicle release from human model neurons (NT2) is sensitive to botulinum toxin A.
Botulinum neurotoxins (BoNTs) internalize into nerve terminals and block the release of neurotransmitters into the synapse. BoNTs are widely used as a therapeutic agent for treatment of movement disorders and recently gained more attention as a biological weapon. Consequently, there is strong interest to develop a cell-based assay platform to screen the toxicity and bioactivity of the BoNTs. In this study, we present an in vitro screening assay for BoNT/A based on differentiated human embryonal carcinoma stem (NT2) cells. The human NT2 cells fully differentiated into mature neurons that display immunoreactivity to cytoskeletal markers (βIII-tubulin and MAP2) and presynaptic proteins (synapsin and synaptotagmin I). We showed that the human NT2 cells undergo a process of exo-endocytotic synaptic vesicle recycling upon depolarization with high K(+) buffer. By employing an antibody directed against light chain of BoNT/A, we detected internalized toxin as a punctate staining along the neurites of the NT2 neurons. Using well-established methods of synaptic vesicle exocytosis assay (luminal synaptotagmin I and FM1-43 imaging) we show that pre-incubation with BoNT/A resulted in a blockade of vesicle release from human NT2 neurons in a dose-dependent manner. Moreover, this blocking effect of BoNT/A was abolished by pre-adsorbing the toxin with neutralizing antibody. In a proof of principle, we demonstrate that our cell culture assay for vesicle release is sensitive to BoNT/A and the activity of BoNT/A can be blocked by specific neutralizing antibodies. Overall our data suggest that human NT2 neurons are suitable for large scale screening of botulinum bioactivity.


Long-lasting effects of neonatal ionizing radiation exposure on spatial memory and anxiety-like behavior.
Neonatal ionizing radiation exposure has been shown to induce a cerebellar cytoarchitecture disarrangement. Since cerebellar abnormalities have been linked to an impairment of behavioral functions, the aim of the present work was to investigate whether exposure of developing rats to ionizing radiations can produce behavioral deficits in the adult. Male Wistar rats were X-irradiated with 5Gy within 48h after birth and were tested in a radial maze and in an open field at 30 and 90 days post irradiation. Irradiated rats showed significative changes in spatial, exploratory, and procedural parameters in the radial maze, as well as a significative decrease in anxiety-like behavior, assessed in the open field. These results suggest that ionizing radiations can induce long-lasting spatial memory and anxiety-related changes. A relationship with radiation-induced cerebellar cytoarchitecture abnormalities supports the hypothesis that cerebellar integrity seems to be critical to achieve spatial performance and emotional behavior establishment.


Heliquinomycin, a new inhibitor of DNA helicase, produced by Streptomyces sp. MJ929-SF2 I. Taxonomy, production, isolation, physico-chemical properties and biological activities.
Heliquinomycin was isolated as a part of a program designed to find inhibitors of DNA helicase from microbial sources. It was purified from the culture broth of Streptomyces sp. MJ929-SF2 by solvent extraction and serial chromatographies of centrifugal partition chromatography, Sephadex LH-20 and Capcell Pak C18 (HPLC). The isolated red powder was analyzed to have the molecular formula of C33H30O17. It inhibited partially purified DNA helicase from HeLa cell in a non-competitive manner with the inhibition constant (Ki) of 6.8 mM. Heliquinomycin exhibited biological activity against microorganisms including MRSA, and cultured cell lines.


Surgery of gastric cancer: treatment of post-operative enterocutaneous fistulas.
Anastomosis dehiscences after gastrectomy are a major post-operative complication often resulting in the death of the patient. 13 patients with dehiscences resulting in enterocutaneous fistulae have been treated with parenteral and/or enteral nutrition. Spontaneous closure rate was 84.6% and mean time for fistula closure was 22.4 days from the onset of nutritional treatment. On the base of nutritional effectiveness no difference could be found between enteral and parenteral nutrition in this series.


The measurement of relative antibody affinity by ELISA using thiocyanate elution.
A variation of the standard ELISA assay was used to determine the relative affinities of six murine monoclonal anti-dinitrophenol (DNP) antibodies for DNP-bovine serum albumin (DNP-BSA). The procedure involved exposing replicate wells containing antibody bound to antigen to increasing concentrations of the chaotropic thiocyanate ion. Resistance to thiocyanate elution was utilized as a measure of affinity and an index (affinity index) representing a 50% reduction in initial absorbance was used to rank the anti-DNP antibodies in order of increasing affinity. For comparison, the affinity constants of the six monoclonals were determined by equilibrium dialysis with 3H-epsilon-DNP-L-lysine as antigen. A significant correlation (P less than 0.02) was observed between the ranking orders obtained using the two methods, thereby demonstrating the applicability of the elution technique for the measurement of relative antibody affinity.


Treatment practices for aspirin-exacerbated respiratory disease: analysis of a national insurance claims database.
Aspirin-exacerbated respiratory disease (AERD) is the triad of asthma, nasal polyposis, and sensitivity to cyclooxygenase-1 inhibitors. Treatment options include medical management, surgical intervention, and aspirin desensitization (AsaD). AERD patients were identified using the MarketScan Database from 2009 to 2015. Patients were included using International Classification of Diseases, 9th edition (ICD-9) codes for asthma, nasal polyposis, and drug allergy. Treatments were determined by Current Procedural Terminology (CPT) codes for drug desensitization and endonasal procedures. Geographic trends and timing of interventions between those exposed and not exposed to desensitization were explored. A total of 5628 patients met inclusion criteria for AERD, with mean age 46 years, 60% female; 395 (7%) underwent AsaD and 2171 (39%) underwent sinus surgery. Among patients who were desensitized, 229 (58%) underwent surgery, of whom 201 (88%) had surgery prior to AsaD (median [quartile 1, quartile 3]; 61 days [30, 208] prior to desensitization). For patients undergoing surgery following AsaD (n = 46), surgery was performed a median of 302 (163, 758) days after AsaD. Nineteen patients had multiple surgeries post-AsaD with median time between surgeries being 734 days (312, 1484); 261 patients were not desensitized to aspirin but did undergo multiple surgeries, with the median of the median time between surgeries being 287 days (15, 617), which is shorter than for patients post-AsaD (p < 0.001). A very small percentage of AERD patients undergo AsaD. Patients who had AsaD underwent surgery approximately 2 months prior to AsaD. Patients who underwent AsaD experienced an increased time between surgeries compared to patients who did not undergo AsaD.


Clinical characteristics and severity of diabetic ketoacidosis: A cross-sectional study from a tertiary hospital in Ghana.
Diabetic ketoacidosis (DKA) is a common, severe and often fatal complication of diabetes. This study aimed to investigate the clinical characteristics and precipitants of DKA, as well as factors associated with DKA severity in Ghanaian patients. Cross-sectional study of the medical records of all 70 adult patients >18 years managed for DKA in the adult emergency room of Korle-Bu Teaching Hospital in Ghana from March 2019 to July 2019. DKA diagnosis was based on hyperglycaemia >11.0 mmol/L, ketonuria (more than 2+) plus acidaemia of (pH < 7.3) or bicarbonate (HCO3 - ) <15.0 mmol/L. However, when serum bicarbonate and pH were not available, clinical signs of acidosis, for example, Kussmaul breathing aided in the diagnosis. DKA severity was assessed based on the Joint British Diabetes Societies (JBDS) guidelines of factors suggestive of severe DKA. Multivariable logistic regression was used to determine the factors associated with DKA severity. Odds ratio and 95% confidence interval for factors associated with DKA severity were determined. The mean (±standard deviation) age, diabetes duration and blood sugar at admission were 44.06 (±16.23) years, 7.19 (±6.04) years and 26.37 (±6.70) mmol/L, respectively. Females comprised 51.4% of the study population. The most common presenting symptoms were generalised weakness (30.0%) and fever (14.3%). The major precipitants were infection (70.0%) and non-compliance (22.9%). Overall, 71.4% of participants had features suggestive of severe DKA. In a multivariable regression model, Type 2 diabetes was associated with over fourfold decreased odds of severe DKA (OR 0.23, 95% CI [0.07-0.76], p = 0.016). Patient education on prevention of DKA was documented for only 18.6% of patients before being discharged. In this study, more than 70% of the study participants had features suggestive of severe DKA, with infection being the most common precipitant of DKA. 51.4% of patients had Type 2 diabetes which was associated with a statistically lower risk of severe DKA. Female sex tended to be positively associated with DKA severity. In a setting where the venous/arterial pH and bicarbonate levels may be inaccessible and/or unaffordable, using clinical features as found in the JBDS guidelines may help categorise patients and escalate care when needed. Indeed it may be useful to validate the use of the JBDS criteria for use in such settings.


Evaluation of a community-based intervention for health and economic empowerment of marginalized women in India.
Empowered women have improved decision-making capacity and can demand equal access to health services. Community-based interventions based on building women's groups for awareness generation on maternal and child health (MCH) are the best and cost-effective approaches in improving their access to health services. The present study evaluated a community-based intervention aimed at improving marginalized women's awareness and utilization of MCH services, and access to livelihood and savings using the peer-led approach from two districts of India. We used peer educators as mediators of knowledge transfer among women and for creating a supportive environment at the household and community levels. The intervention was implemented in two marginalized districts of Uttar Pradesh, namely Banda and Kaushambi. Two development blocks in each of the two districts were selected randomly, and 24 villages in each of the four blocks were selected based on the high percentage of a marginalized population. The evaluation of the intervention involved a non-experimental, 'post-test analysis of the project group' research design, in a mixed-method approach. Data were collected at two points in time, including qualitative interviews at the end line and tracking data of the intervention population (n = 37,324) through an online management information system. Most of the women in Banda (90%) and Kaushambi (85%) attended at least 60% of the education sessions. Around 39% of women in Banda and 35% of women in Kaushambi registered for the livelihood scheme, and 94 and 80% of them had worked under the scheme in these two places, respectively. Women's awareness about MCH seemed to have increased post-intervention. The money earned after getting work under the livelihood scheme or from daily savings was deposited in the bank account by the women. These savings helped the women investing money at times of need, such as starting their work, in emergencies for the medical treatment of their family members, education of their children, etc. CONCLUSION: Peer-led model of intervention can be explored to improve the combined health and economic outcomes of marginalized women.


Incidence, Management, and Outcomes of Very Early Onset Inflammatory Bowel Diseases and Infantile-Onset Disease: An Epi-IIRN Study.
In this nationwide study from the Israeli Inflammatory Bowel Disease Research Nucleus, we aimed to describe the incidence of very early onset inflammatory bowel diseases (VEOIBDs) with a focus on infantile-onset disease and to compare management and disease course with older children. Data were retrieved from the 4 Israeli Health Maintenance Organizations covering 98% of the population. Pediatric-onset IBD was categorized as follows: adolescent onset (10 to <18 y), early onset (6 to <10 y), VEOIBD (0 to <6 y), toddler onset (2 to <6 y), and infantile onset (<2 y). A total of 5243 children with 35,469 person-years of follow-up evaluation, were diagnosed with IBD during 2005 to 2020: 4444 (85%) with adolescent onset, 548 (10%) with early onset, and 251 (4.8%) with VEOIBD, of whom 81 (1.5%) had infantile onset. The incidence of pediatric-onset IBD increased from 10.8 per 100,000 in 2005 to 15.3 per 100,000 in 2019 (average annual percentage change, 2.8%; 95% CI, 2.2%-3.4%), but that of VEOIBD remained stable (average annual percentage change, 0%; 95% CI, -2.5% to 2.6%). The infantile-onset and toddler-onset groups were treated less often with biologics (36% and 35%, respectively) vs the early onset (57%) and adolescent-onset groups (53%; P < .001). The time to steroid dependency was shorter in infantile-onset (hazard ratio [HR], 2.1; 95% CI, 1.5-2.9) and toddler-onset disease (HR, 1.6; 95% CI, 1.2-2.0) vs early onset and adolescent-onset disease, but time to hospitalizations, time to surgery, and growth delay were worse only in infantile-onset disease. In a multivariable model, infantile-onset patients had a higher risk for surgery (HR, 1.4; 95% CI, 1.1-1.9) and hospitalization (HR, 1.7; 95% CI, 1.2-2.4) than the toddler-onset group. The incidence of VEOIBD remained stable. Infantile-onset IBD had worse outcomes than older children, while toddler onset had mostly similar outcomes, despite less frequent use of biologics.


Does the 'Educational Alliance' conceptualize the student - supervisor relationship when conducting a master thesis in medicine? An interview study.
Completing a master thesis (MT) is mandatory in many undergraduate curricula in medicine but a specific educational framework to guide the supervisor-student relationship during the MT has not been published. This could be helpful to facilitate the MT process and to more effectively reach the learning objectives related to science education in medicine. An attractive model for this purpose is the 'Educational Alliance' (EA), which focusses on the three components 'clarity and agreement on (a) goals, (b) tasks and (c) relationship & roles'. This study investigated factors that can either facilitate or hinder the process of MTs, and related these to the components of the EA. We conducted semi-structured face-to-face interviews with 20 students and - separately - with their 20 corresponding supervisors, after the MT had been accepted. The interviews included open questions on factors facilitating or hindering the success of the MT. Audio recordings of the interviews were anonymized and transcribed, and then analysed by qualitative content analysis. Also, quantitative data were gathered on satisfaction with the MT process and the supervisory quality (using Likert-type questions). We were able to analyse all 40 interviews, related to 20 MTs. From the transcripts, we extracted 469 comments related to the research question and categorized these into the four main categories (a) 'Preparation', (b) 'Process', (c) 'Atmosphere', (d) 'Value of the MT'. Interviewees highlighted the importance of a careful preparation phase, clear expectations, a clear research plan, thorough and timely feedback, mutual agreement on timelines, and a positive working atmosphere. Each of these factors could be brought in line with the three components of the EA framework: agreement and clarity of goals, tasks, relationships & roles. Satisfaction with the MT process was rated 8.75 ± 1.22 SD (of 10) points by supervisors, and 7.80 ± 1.61 SD points by students, while supervision quality was rated + 1.51 ± 0.63 SD (scale from - 2 to + 2) by supervisors, and + 1.26 ± 0.93 SD by students. We propose the EA framework as a useful guidance for students, supervisors, and the university towards conducting successful MTs in medicine. Based on the findings, we provide specific recommendations for students, supervisors, and university.


Electromyographic assessment of ulnar nerve motor block induced by lidocaine.
To determine the differences in the onset time and duration of motor block produced by lidocaine 1% and lidocaine 2% via a quantitative and objective method, the measurement of compound muscle action potentials (CMAPs). Prospective study. Main operating rooms of a university hospital. 20 consecutive patients undergoing surgery not requiring intraoperative muscle relaxation. General anesthesia with unilateral ulnar nerve block was administered. In patients' nondominant (experimental) arms, an insulated block needle was placed adjacent to the ulnar nerve at the wrist while continuous nerve stimulation was delivered to ensure its proper placement. Through this needle, lidocaine 1% or lidocaine 2% was injected. The dominant (control) arm received no injection. Monitoring of ulnar nerve-evoked CMAPs was performed simultaneously on both arms. Ulnar nerve function was assessed at baseline and then at 10-second intervals by automatically measuring the amplitude of the evoked CMAPs on a two-channel electromyogram. The mean (+/- SEM) baseline CMAP amplitude prior to injection of lidocaine 1% was 3.10 +/- 0.87 mV and 3.06 +/- 0.89 mV for the experimental and control ulnar nerves, respectively (p = NS); for lidocaine 2%, baseline CMAP amplitude was 3.58 +/- 1.39 mV and 3.70 +/- 1.46 mV, respectively (p = NS). Over the course of the study, the control CMAP amplitude varied by < 12%. At the experimental ulnar nerve, 90% CMAP decrease after injection of lidocaine 1% and lidocaine 2% occurred 7.5 +/- 2 minutes and 5 +/- 1.5 minutes, respectively (p = NS), whereas maximal blockade was achieved after 15 +/- 3 minutes and 11 +/- 5 minutes, respectively (p = NS). Recovery of CMAP to 90% of baseline occurred 184 +/- 31 minutes after injection of lidocaine 1% and 248 +/- 30 minutes following lidocaine 2% (p = NS). The present study describes a technique that can be used in vivo to objectively measure the speed of onset and duration of local anesthetic-induced motor blockade. Although statistically not different, lidocaine 2% demonstrated a faster onset and longer duration of ulnar nerve motor block than lidocaine 1%.


Interaction of morphine, etorphine and enkephalins with dopamine-stimulated adenylate cyclase of monkey amygdala.
Adenylate cyclase activity (AC) of homogenates of monkey amygdaloid nucleus was approximately doubled in the presence of dopamine (10 micrometer). Morphine, etorphine, and several enkephalin analogs (met-enkephalin, D-ala2-met-enkephalin, and D-met2, pro5-enkephalinamide) were capable of inhibiting the stimulation of AC produced by dopamine (90-100% with etorphine or D-ala2-met-enkephalin). Unlike morphine and etorphine, the peptides exhibited bell-shaped dose-response curves for this inhibition with maximal effects at approximately 1 X 10(-7) M, but negligible effects at 1 X 10(-5) M. Under the conditions studied, only etorphine inhibited basal AC. Naloxone antagonized the inhibitory effects of each of the opioids tested, and dextrorphan, an inactive L-(+)-opiate, failed to inhibit the dopamine response. Together these data indicate that the effects were mediated via the classically described stereospecific opiate receptor. The relative order of potency (etorphine greater than enkephalins greater than morphine) was similar to that previously reported for the binding affinities of these drugs in rat brain homogenates. The influence of narcotic agents on dopamine stimulated AC was eliminated by either freezing the amygdaloid tissue or preincubating the homogenate at 4 degrees C; the dopamine responses, however, could still be elicited. The narcotic receptor interaction with the adenylate cyclase thus appears to be distinct from and more labile than that of the dopamine receptor. Gpp(NH)p-stimulated AC was not inhibited by morphine. It is postulated that the inhibition involves interaction of opiate receptors with catalytic units of dopamine-stimulated AC, but not with other cyclase species which may provide the major component of Gpp(NH)p-stimulated activity in amygdala.


Treatment of injuries to the ulnar side of the wrist occurring with distal radial fractures.
We still do not know how to best treat the DRUJ condition that was recognized 200 years ago by Abraham Colles and later addressed in Frykman's classic thesis. To improve the outcome, we must recognize the differences be-tween osteoporotic and other fractures and understand the importance of ligament injuries,especially in patients under the osteoporotic age. However, our current problem is that neither the initial ligament injury nor the posttraumatic laxity is detectable with radiographic methods,which creates future challenges regarding diagnosis and treatment. We therefore have to critically analyze each fracture in each patient and be aware of the complexity of the entire injury to the wrist.


Fast and Frictionless: A Novel Approach to Radiology Appointment Scheduling Using a Mobile App and Recommendation Engine.
Many outpatient radiology orders are never scheduled, which can result in adverse outcomes. Digital appointment self-scheduling provides convenience, but utilization has been low. The purpose of this study was to develop a "frictionless" scheduling tool and evaluate the impact on utilization. The existing institutional radiology scheduling app was configured to accommodate a frictionless workflow. A recommendation engine used patient residence, past and future appointment data to generate three optimal appointment suggestions. For eligible frictionless orders, recommendations were sent in a text message. Other orders received either a text message for the non-frictionless app scheduling approach or a call-to-schedule text. Scheduling rates by type of text message and scheduling workflow were analyzed. Baseline data for a 3-month period prior to the launch of frictionless scheduling showed that 17% of orders that received an order notification text were scheduled using the app. In an 11-month period after the launch of frictionless scheduling, the rate of app scheduling was greater for orders that received a text message with recommendations (frictionless approach) versus app schedulable orders that received a text without recommendations (29% vs. 14%, p < 0.01). Thirty-nine percent of the orders that received a frictionless text and scheduled using the app used a recommendation. The most common recommendation rules chosen for scheduling included location preference of prior appointments (52%). Among appointments that were scheduled using a day or time preference, 64% were based on a rule using the time of the day. This study showed that frictionless scheduling was associated with an increased rate of app scheduling.


Incorporating utility-weights when comparing two diagnostic systems: a preliminary assessment.
We sought to develop a new index that incorporates utility-weights when assessing the overall performance of a diagnostic system and to provide a statistical test for comparing two indices in a paired study design. The area under the receiver operating characteristic (ROC) curve (AUC) was used as the basis for constructing a new index. The index we propose represents a weighted average of class-specific AUCs each of which relates to a class of pairs of actually negative (normal) and actually positive (abnormal) cases with a specific predetermined utility (or clinical importance). For each pair of normal-abnormal cases, the utility is defined a priori and based on external (covariate) information. In the proposed approach utility-weights represent the relative importance (utility) of discriminating between different types of normal and abnormal cases (pairs of the same type are combined in the classes termed utility-classes). We also describe a simple nonparametric procedure for comparing the proposed indices as computed from paired data. Computer simulations were conducted to evaluate the behavior of the type I error of the proposed test in the simple albeit important instance of two utility-classes. The new index provides an extension of the commonly used area under the ROC curve. It allows for incorporation of utility-weights into the analysis and reduces to the conventional AUC index when all assigned utility-weights are equal to unity. Computer simulations indicate that in the considered scenario of two utility-classes, the type I error of the proposed test is comparable to that of the conventional nonparametric test for equality of AUC indices. The proposed index and the statistical test provide a practical approach of incorporating utilities when comparing diagnostic systems.


Tentacles of in vitro-grown round-leaf sundew (Drosera rotundifolia L.) show induction of chitinase activity upon mimicking the presence of prey.
Induction of plant-derived chitinases in the leaves of a carnivorous plant was demonstrated using aseptically grown round-leaf sundew (Drosera rotundifolia L.). The presence of insect prey was mimicked by placing the chemical inducers gelatine, salicylic acid and crustacean chitin on leaves. In addition, mechanical stirring of tentacles was performed. Chitinase activity was markedly increased in leaf exudates upon application of notably chitin. Application of gelatine increased the proteolytic activity of leaf exudates, indicating that the reaction of sundew leaves depends on the molecular nature of the inducer applied. In situ hybridization of sundew leaves with a Drosera chitinase probe showed chitinase gene expression in different cell types of non-treated leaves, but not in the secretory cells of the glandular heads. Upon induction, chitinase mRNA was also present in the secretory cells of the sundew leaf. The combined results indicate that chitinase is likely to be involved in the decomposition of insect prey by carnivorous plants. This adds a novel role to the already broad function of chitinases in the plant kingdom and may contribute to our understanding of the molecular mechanisms behind the ecological success of carnivorous plants in nutritionally poor environments.


Membrane associated particles of the presynaptic active zone in rat spinal cord. A morphometric analysis.
The distribution of membrane-associated particles in the presynaptic membrane was investigated in the spinal cord of unanesthetized and anesthetized rats by freeze etching electron microscopy. Both 'external' face (EF) and 'protoplasmic' face (PF) were examined. Particles were classified according to size in two categories: small particles (diameter 5.0-8.7 nm) and large particles (diameter 8.7-13.7 nm). The presynaptic region was subdivided into an active and a surrounding zone, depending on specific ultrastructural criteria. The density of large particles in the PF was found to be significantly higher in the active as compared with the surrounding zone in both unanesthetized and anesthetized rats. Thus, the presence of large particles represents an important feature of the active zone. Considerably more large particles were found in the waking than in the barbiturized state. This difference is paralleled by a vast increase of vesicle attachment sites in the presynaptic membrane of unanesthetized animals reported by Streit et al. and confirmed in the present study. It is suggested that the two phenomena could be interrelated and that the large particles may represent calcium channels and thus provide the morphological substrate for the mechanism of excitation-secretion coupling.


[Malignant hyperthermia. III. Progress report on serum CK activity, questionable screening test, effect of dantrolene in family members with a hereditary disposition].
18 members of a MH prone family have been studied to determine the value of various clinical methods which might be helpful in detecting the disposition. Tests included serum levels of CK over a period of 3 years, and of potassium, the influences of muscle provocation and pretreatment with dantrolene, concentration of cholesterol in erythrocytes and cholinesterase (activity and genotype). The results show relative constant measurements of the enzyme; there was, however, no evidence of any other reliable method in assessing the disposition.


Thioredoxin Interacting Protein Is Required for a Chronic Energy-Rich Diet to Promote Intestinal Fructose Absorption.
Increased consumption of fats and added sugars has been associated with an increase in metabolic syndromes. Here we show that mice chronically fed an energy-rich diet (ERD) with high fat and moderate sucrose have enhanced the absorption of a gastrointestinal fructose load, and this required expression of the arrestin domain protein Txnip in the intestinal epithelial cells. ERD feeding induced gene and protein expression of Glut5, and this required the expression of Txnip. Furthermore, Txnip interacted with Rab11a, a small GTPase that facilitates the apical localization of Glut5. We also demonstrate that ERD promoted Txnip/Glut5 complexes in the apical intestinal epithelial cell. Our findings demonstrate that ERD facilitates fructose absorption through a Txnip-dependent mechanism in the intestinal epithelial cell, suggesting that increased fructose absorption could potentially provide a mechanism for worsening of metabolic syndromes in the setting of a chronic ERD.


The role of vitamin B12 and folate in carcinogenesis.
The roles of vitamin B12 and folate in carcinogenesis are largely extensions of and linked to their roles in normal metabolism, particularly 1-carbon unit metabolism. A possible key area may be hypomethylation to "switch on" genes and methylation to "switch them off." Some vitamin analogues may act as antivitamins in these reactions, as may some vitamin-binding proteins. Others may act as specific delivery proteins. Using appropriate radioactive substrates and suspensions of vitamin-dependent normal and malignant cells, it may be possible to work out their positive and negative control of DNA synthesis.


Avian colonic ion transport: effects of corticosterone and dexamethasone.
1. Corticosterone, a natural corticosteroid hormone in birds, when injected into domestic fowl (Gallus domesticus) (2000 micrograms.kg-1, 4-5 h before experiment) increases both the basal Isc (short-circuit current) and amiloride-sensitive Isc as well as the PD across the colon in vitro. Dexamethasone, a synthetic analogue (650 micrograms.kg-1, 4-5 h before experiment) also increases the basal and amiloride-sensitive Isc as well as PD in these preparations. 2. In marked contrast, longer term injection or infusion of dexamethasone (650 micrograms.kg-1) for 3 or more days caused a decline in basal Isc and PD (the PD often reversed with the serosal side becoming electronegative) and a drop in resistance. However in these preparations, the amiloride-sensitive Isc was significantly elevated which could be accounted for by an increase in net Na flux. 3. No significant change occurs in net flux of Cl or K although unidirectional fluxes in both directions were increased for both ions in birds given dexamethasone for 3 days. 4. A disparity between the basal Isc and the amiloride-sensitive Isc appeared in these preparations from dexamethasone injected birds reflecting the transport of other ions, possibly HCO3- or H+. The possible role of corticosterone in mineral metabolism of birds is discussed.


The RNA N-glycosidase activity of ricin A-chain. The characteristics of the enzymatic activity of ricin A-chain with ribosomes and with rRNA.
Ricin A-chain cleaves the N-glycosidic bond at A-4324 in 28 S rRNA when intact rat ribosomes are the substrate. Cleavage occurs at a concentration of the toxin of 1 X 10(-10) M, and specificity for this single residue is retained when the concentration is as high as 3 X 10(-7) M. The apparent Michaelis constant (Km) for the reaction is 2.6 microM, and the turnover number (Kcat) is 1777 min-1. The same N-glycosidic bond is cleaved by ricin A-chain in naked 28 S rRNA, but at a greatly reduced rate. The Km value for this reaction is 5.8 microM. The results suggest that the A-chain has a similar affinity for 28 S rRNA in ribosomes and in the absence of ribosomal proteins. Ricin A-chain has no effect on 23 S rRNA in Escherichia coli ribosomes, however, the N-glycosidic bond at A-2600 in naked 23 S rRNA is cleaved by the toxin; this corresponds to the ricin site in eukaryotic 28 S rRNA. Since the Km value (3.3 microM) for the reaction with E. coli 23 S rRNA approximates that obtained with rat liver ribosomes, it is possible that E. coli ribosomal protein(s) protect this site against ricin attack in intact ribosomes. Ricin A-chain also acted on naked 16 S rRNA cleaving the N-glycosidic bond of adenine at position 1014. The results suggest that ricin A-chain recognizes a specific structure in rRNA, perhaps a loop and stem having the sequence GAGA in the loop.