Sessile ileum, subhepatic cecum, and uncinate appendix that might lead to a diagnostic dilemma.
The subhepatic position of the cecum and appendix is a result of embryological reasons. Subhepatic appendicitis can cause diagnostic dilemmas. During the dissection of an adult male cadaver aged approximately 70 years, the subhepatic position of the cecum and appendix was noted. The appendix made a "U"-shaped bend and its tip was located in the paracolic position. The cecum had appendices epiploicae, and the terminal part of the ileum was retroperitoneal and had ascended vertically to the cecum from the right iliac fossa. Functionally, the sessile part of the ileum might restrict its peristaltic movements. The abnormal position of the terminal ileum might be mistaken for an ascending colon during laparoscopic surgery. The subhepatic position of the cecum and appendix might cause confusion in the diagnosis of acute appendicitis because the tenderness in such cases is not located at the McBurney's point.


DNA array profiling of gene expression changes during maize embryo development.
We are using DNA microarray-based gene expression profiling to classify temporal patterns of gene expression during the development of maize embryos, to understand mRNA-level control of embryogenesis and to dissect metabolic pathways and their interactions in the maize embryo. Genes involved in carbohydrate, fatty acid, and amino acid metabolism, the tricarboxylic acid (TCA) cycle, glycolysis, the pentose phosphate pathway, embryogenesis, membrane transport, signal transduction, cofactor biosynthesis, photosynthesis, oxidative phosphorylation and electron transfer, as well as 600 random complementary DNA (cDNA) clones from maize embryos, were arrayed on glass slides. DNA arrays were hybridized with fluorescent dye-labeled cDNA probes synthesized from kernel and embryo poly(A)(+)RNA from different stages of maize seed development. Several characteristic developmental patterns of expression were identified and correlated with gene function. Patterns of coordinated gene expression in the TCA cycle and glycolysis were analyzed in detail. The steady state level of poly(A)(+) RNA for many genes varies dramatically during maize embryo development. Expression patterns of genes coding for enzymes of fatty acid biosynthesis and glycolysis are coordinately regulated during development. Genes of unknown function may by assigned a hypothetical role based on their patterns of expression resembling well characterized genes. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s10142-002-0046-6.


Oxidative stress in songbirds exposed to dietary methylmercury.
Long-term, sublethal methylmercury exposure can cause reproductive depression, immune suppression, endocrine disruption and other problems in birds. We used two biomarkers to detect oxidative stress in livers of zebra finches (Taeniopygia guttata) developmentally exposed to sublethal levels of dietary methylmercury (0.0, 0.3, 0.6, 1.2, or 2.4 μg/g wet weight in diet). Our findings indicate that young adult finches exposed to environmentally relevant concentrations of mercury in ovo and through their diets, exhibited oxidative stress in their livers. We measured the ratio of the antioxidant glutathione in its reduced form (GSH) versus its oxidized form (GSSG) and the activity of the superoxide dismutase (SOD) enzyme suite. Blood total mercury served as a proxy for liver mercury concentration, and was on average 8.4 times the dietary dose (e.g., birds consuming 0.6 μg/g had blood mercury levels of ~5 μg/g on a wet weight basis). Consistent with what is known from large, aquatic bird species, there was a significant, negative relationship between GSH/GSSG ratios and tissue mercury concentrations, which is indicative of oxidative stress. This relationship was driven by a significant increase in the oxidized glutathione in the livers of birds with higher blood mercury levels. SOD activity was also found to have a significant, negative relationship with blood mercury.


Outcomes Following Bronchial Artery Embolisation for Haemoptysis in Cystic Fibrosis.
Bronchial artery embolisation (BAE) is recommended for the treatment of massive haemoptysis in cystic fibrosis (CF), but there are no randomised controlled trials of this therapy and its role in sub-massive haemoptysis is unclear. This study aimed to determine the outcomes and safety of BAE in adults with CF. All patients with CF undergoing BAE at our centre between March 2011 and January 2015 were identified at the time of the procedure. Patient records were reviewed at hospital discharge, death or one month post-procedure (whichever was soonest). Follow-up continued to January 2016. Severity of haemoptysis was classified as: massive (>240 ml/24 h or >100 ml/day for ≥2 days), moderate-severe (>20 ml/24 h) or mild (<20 ml/24 h). Twenty-seven patients underwent 51 BAE procedures over a median follow-up period of 26 months (range 1-54). Ten patients (37%) required more than one BAE during the study. BAE was performed for massive haemoptysis in 18 cases (35%). Haemoptysis recurred after 31 (61%) of BAE procedures with no difference in recurrence rates between massive and sub-massive haemoptysis. Side effects were reported after 61% of procedures with chest pain the most common adverse event . Mortality after first BAE in the study was 3.9% at 30 days and 14.8% at 12 months. No significant predictors of mortality were identified. BAE is often effective in controlling haemoptysis but is associated with considerable morbidity and high recurrence rates.


A traumatic tale of two cities: a comparison of outcomes for adults with major trauma who present to differing trauma centres in neighbouring Canadian provinces.
While the use of formal trauma teams is widely promoted, the literature is not clear that this structure provides improved outcomes over emergency physician delivered trauma care. The goal of this investigation was to examine if a trauma team model with a formalized, specialty-based trauma team, with specific activation criteria and staff composition, performs differently than an emergency physician delivered model. Our primary outcome was survival to discharge or 30 days. An observational registry-based study using aggregate data from both the New Brunswick and Nova Scotia trauma registries was performed with data from April 1, 2011 to March 31, 2013. Inclusion criteria included patients 16 years-old and older who had an Injury Severity Score greater than 12, who suffered a kinetic injury and arrived with signs of life to a level-1 trauma centre. 266 patients from the trauma team model and 111 from the emergency physician model were compared. No difference was found in the primary outcome of proportion of survival to discharge or 30 days between the two systems (0.88, n=266 vs. 0.89, n=111; p=0.8608). We were unable to detect any difference in survival between a trauma team and an emergency physician delivered model.


Vaccinia virus E3L protein is an inhibitor of the interferon (i.f.n.)-induced 2-5A synthetase enzyme.
Induction of apoptosis in mammalian cells by double-stranded (ds) RNA-dependent enzymes, protein kinase (PKR), and 2-5A-synthetase/RNase L (referred to as the 2-5A system) might be a mechanism mediating anticellular and antiviral actions of interferon (i.f.n.). To counteract the effect of i.f.n., animal viruses have acquired genes that block specific i.f.n. pathways. Among poxviruses, vaccinia virus (VV) encodes E3L, a dsRNA-binding protein, which inhibits activation of i.f.n.-induced PKR. It has been proposed that E3L might also block activation of the 2-5A system, but direct proof is lacking. To establish if E3L inhibits the 2-5A system, we have developed a method to assay apoptosis induced by increased production of enzymes in the 2-5A pathway, as well as of their putative modulators. This assay is based on the use of cells derived from homozygous PKR knockout mice (Pkr-/-) infected with a VV mutant lacking E3L (delta E3L) and transiently transfected with a luciferase reporter gene together with plasmid vectors expressing 2-5A-synthetase, RNase L, or E3L, all controlled by the same inducible promoter. We found that expression of 2-5A-synthetase inhibited luciferase activity in a dose-response manner, reaching inhibition values of 80% relative to transfections with control plasmids. Similar results were obtained by transfection with an RNase L vector, although in this case the extent of inhibition was further enhanced upon coexpression of 2-5A-synthetase and RNase L. Inhibition of protein synthesis mediated by the 2-5A system correlated well with induction of apoptosis. Transfection of cells with a plasmid vector expressing E3L together with 2-5A-synthetase completely prevented apoptosis induced by this enzyme. We conclude that VV E3L acts as an inhibitor of the i.f.n.-induced 2-5A-synthetase enzyme.


Pseudophakic retinal detachment.
In 40 cases of pseudophakic retinal detachment 88% of the retinas were reattached, and 20% of the patients had a final visual acuity of 6/12 or better. Poor visualization of the fundus in 38% of the cases and the inability to find retinal breaks in 20% of the cases were major problems in management.


Agenesis of the dorsal pancreas associated with mucinous cysts and chronic calcific non-alcoholic pancreatitis.
Agenesis of the dorsal pancreas is a rare malformation. Since 1911 and until 2008, 107 cases have been reported. Herein we report a new case on agenesis of the dorsal pancreas associated with mucinous is and chronic calcific non-alcoholic pancreatitis.


Orthotic collar in the treatment of intertriginous impetigo.
This article illustrates a manner in which physical therapy can provide adjunctive treatment management of an infant with intertriginous impetigo. This treatment program was an interdisciplinary approach to a difficult clinical problem. The medical staff concluded that implementation of the cervical orthosis expedited the patient's recovery and reduced the potential complications of the disease process.


Dose dense cyclophosphamide, methotrexate, fluorouracil is feasible at 14-day intervals: a pilot study of every-14-day dosing as adjuvant therapy for breast cancer.
Cyclophosphamide/methotrexate/fluorouracil (CMF) is a proven adjuvant option for patients with early-stage breast cancer. Randomized trials with other regimens demonstrate that dose-dense (DD) scheduling can offer greater efficacy. We investigated the feasibility of administering CMF using a DD schedule. Thirty-eight patients with early-stage breast cancer were accrued from March 2008 through June 2008. They were treated every 14 days with C 600, M 40, F 600 (all mg/m2) with PEG-filgrastim (Neulasta®) support on day 2 of each cycle. The primary endpoint was tolerability using a Simon's 2-stage optimal design. The design would effectively discriminate between true tolerability (as protocol-defined) rates of ≤ 60% and ≥ 80%. The median age was 52-years-old (range, 38-78 years of age). Twenty-nine of the 38 patients completed 8 cycles of CMF at 14-day intervals. Dose-dense adjuvant CMF is tolerable and feasible at 14-day intervals with PEG-filgrastim support.


Use of the triolein breath test for the demonstration of fat malabsorption in coeliac disease.
Different screening tests for fat malabsorption were evaluated in patients with coeliac disease. The triolein breath test correlated well with results of faecal fat determination. Urinary excretion of oxalate with or without dietary oxalate loading was not correlated with faecal fat measurements. It is concluded that the triolein breath test is a useful procedure for the detection of fat malabsorption in patients with coeliac disease.


The Prospective Influence of Trait Alexithymia on Intrusive Memories: What Is the Role of Emotional Recognition Memory?
Posttraumatic stress disorder (PTSD) is often considered to be a disorder of memory as patients suffer from fragmented uncontrollable memories (intrusions) whilst experiencing difficulties in intentionally retrieving details of the traumatic event. Recent research suggests that trait-related deficits in the identification of emotional states (alexithymia) may impact emotional memory processes in a way that promotes intrusion formation in PTSD. Therefore, we investigated the influence of alexithymia on intrusive re-experiencing and emotional recognition memory in a prospective analog study. Twenty-six healthy participants took part in a laboratory experiment, which combined two independent paradigms. Participants were exposed to a traumatic film (first session) and completed an episodic memory task comprising neutral and emotional stimuli (second session). In between sessions, participants recorded intrusive memories of the film. Individuals with higher trait alexithymia (HTA) reported an increased number of intrusions on the day of film presentation. Moreover, analyses of memory performance revealed a negative correlation between alexithymia and emotional recognition memory. Further analyses suggest that reduced emotional recognition memory, as evident in individuals with HTA, may, in turn, be associated with enhanced intrusive re-experiencing. As such, the current findings provide first indications regarding the role of alexithymia in emotional learning and PTSD. Future studies should further investigate these associations as well as potential implications for the treatment of PTSD.


Specific and common antigens of Trichomonas vaginalis detected by monoclonal antibodies.
Monoclonal antibodies to Trichomonas vaginalis were prepared by immunizing mice with a cloned isolate of T. vaginalis. Eight antibodies reacted with the same four isolates or strains but did not react with the other T. vaginalis strains or isolates tested. All eight antibodies reacted uniformly with both the body and flagella of T. vaginalis in the immunofluorescence assay but were unreactive by immunoblotting. The antigen(s) recognized by these antibodies was determined to be present on the surface membrane by indirect immunofluorescence assay of live organisms. The antigen(s) was found to be sensitive to periodate oxidation but resistant to pronase digestion. In addition, one monoclonal antibody was derived which reacted with all T. vaginalis isolates or strains tested, as well as with Trichomonas gallinae, Tritrichomonas foetus, and Giardia lamblia. This antibody reacted with the body but not the flagella of Formalin-fixed protozoa in the immunofluorescence assay but failed to react with live organisms. The antigen was found to be periodate resistant but pronase labile. In the immunoblot assay, this antibody detected a single T. vaginalis polypeptide with a molecular weight of 62,000.


Dermatology outpatient clinic outcomes after COVID-19 outbreak: What is new normal?
Coronavirus disease-2019 (COVID-19), which emerged in late 2019 and caused a pandemic, has significantly affected outpatient admissions to dermatology outpatient clinics. There have been changes in the number and composition of the patients who applied to the outpatient clinics. The dermatology outpatient clinic applications have dramatically decreased due to restrictions and prohibitions, and active participation of dermatologists in the field immediately after the pandemic. The composition of the diagnoses has also altered for reasons such as excessive use of hygiene products and types of protective equipment usage related to COVID-19. Intensive precautions have been taken in the first 3 months of the pandemic (March, April, and May). As of 12 May, controlled socialization started with new regulations. This period has been called "the normalization process." This study aims to evaluate the changes of the patients admitted to dermatology outpatient clinics within the normalization process. Despite the increasing number of COVID-19 patients and related deaths in the whole country with the new normal, the admissions to dermatology outpatient clinics have increased. During this period, acne and related diseases, pigmentation disorders, and viral skin infections had increased; dermatoses, xerosis cutis, and superficial fungal infections had reduced. It seems that nonurgent dermatological complaints affect the quality of life of patients and cause the need for an application. Although restrictions reduce these numbers, measures should be taken to protect patients and society during the ongoing pandemic.


Analysis of simple sequence repeats (SSRs) in wild barley from the Fertile Crescent: associations with ecology, geography and flowering time.
Wild barley, Hordeum spontaneum C. Koch, is the progenitor of cultivated barley, Hordeum vulgare. The centre of diversity is in the Fertile Crescent of the Near East, where wild barley grows in a wide range of conditions (temperature, water availability, day length, etc.). The genetic diversity of 39 wild barley genotypes collected from Israel, Turkey and Iran was studied with 33 SSRs of known map location. Analysis of molecular variance (AMOVA) was performed to partition the genetic variation present within from the variation between the three countries of origin. Using classification tree analysis, two (or three) specific SSRs were identified which could correctly classify most of the wild barley genotypes according to country of origin. Associations of SSR variation with flowering time and adaptation to site-of-origin ecology and geography were investigated by two contrasting statistical approaches, linear regression based on SSR length variation and linear regression based on SSR allele class differences. A number of SSRs were significantly associated with flowering time under four different growing regimes (short days, long days, unvernalised and vernalised). Most of the associations observed could be accounted for by close linkage of the SSR loci to earliness per se genes. No associations were found with photoperiodic and vernalisation response genes known to control flowering in cultivated barley suggesting that different genetic factors may be active in wild barley. Novel genomic regions controlling flowering time in wild barley were detected on chromosomes 1HS, 2HL, 3HS and 4HS. Associations of SSRs with site-of-origin ecological and geographic data were found primarily in genomic regions determining plant development. This study shows that the analyses of SSR variation by allele class and repeat length are complementary, and that some SSRs are not necessarily selectively neutral.


Laxative utilization over time in chronic pain patients with opioid-induced constipation.
To determine laxative utilization over time among chronic noncancer pain patients with opioid-induced constipation (OIC). A prospective longitudinal study conducted in the USA, Canada, Germany and UK. Patients on daily opioid therapy for treatment of chronic noncancer pain with OIC were recruited from clinics to complete a survey at Baseline and weeks 2, 4, 6, 8, 12, 16, 20 and 24. 489 patients completed baseline with 452 completing one or more follow-up visits. 128 (28%) were nonlaxative users, 112 (25%) were insufficient laxative users and 212 (47%) were sufficient laxative users. The consistent sufficient laxative users reported the most bowel movements per week. The majority of OIC patients do not take or only intermittently take laxatives.


The Use of Social Media to Combat Research-Isolation.
Research-isolation is a common problem affecting many researchers who are disconnected from their research communities. It can be caused by a number of factors, including physical isolation, unfamiliar research topics, diversity, and the nature of the supervisory relationship. All of these aspects can have an impact on both work and the mental health of researchers. Increasingly, researchers are turning to social media for support, by both looking for communities and for increasing the impact of their work. In this paper, we set out a brief introduction to a range of social media platforms used by researchers and present a discussion of the networks within those platforms aimed at reducing research-isolation. These examples highlight just a few of the number of small communities that have grown online to meet the needs of those seeking support through social media. We conclude with some recommendations for those affected by research-isolation and highlight the need for more research into the role of social media on mental health in academics.


Association of lysyl oxidase-like 1 gene common sequence variants in Greek patients with pseudoexfoliation syndrome and pseudoexfoliation glaucoma.
Three common sequence variants in the lysyl oxidase-like 1 (LOXL1) gene were recently associated with pseudoexfoliation (PEX) and pseudoexfoliation glaucoma (PEXG) in populations from various parts of the world. In this study, the genetic association of these variants was investigated in Greek patients with PEX and PEXG. The three LOXL1 single nucleotide polymorphisms (SNPs), one intronic (rs2165241) and two nonsynonymous coding SNPs (rs1048661: R141L and rs3825942: G153D), were genotyped in a total of 48 unrelated patients with PEX, 35 patients with PEXG, and 52 healthy subjects who had normal findings in repeated ophthalmic examinations. A genetic association study was performed. Between the two coding SNPs, R141L did not show an association with PEX (p=0.297 for allele G, p=0.339 for genotype GG), whereas allele G of G153D showed a significant association (odds ratio [OR]=3.52, 95% confidence interval [CI]=1.735-7.166, p=3.24×10(-4) for allele G, p=0.004 for genotype GG). Likewise, for the intronic SNP of rs2165241, genotype TT (p=0.005) and its corresponding allele T (OR=2.99, 95% CI=1.625-5.527, p=3.53×10(-4)) showed a significant association with PEX. The allele G of G153D showed a significant association with PEXG (OR=3.74, 95% CI=1.670-8.387, p=0.001). The combined haplotype GGT, consisting of all three risk alleles, was associated with PEX (p=0.037), conferring a 1.8-fold of increased risk to the disease (OR=1.799, 95% CI=1.04-3.13). Furthermore, the haplotype GGT presented in 39.8% of the patients with PEX and 26.9% of the controls. Certain genetic variants in LOXL1 confer risk for PEX in Greek populations, confirming in part findings in patients from Northern Europe.


A dot that went for a walk: People prefer lines drawn with human-like kinematics.
A dominant theory of embodied aesthetic experience (Freedberg & Gallese, 2007, Trends in Cognitive Sciences, 11, 197) posits that the appreciation of visual art is linked to the artist's movements when creating the artwork, yet a direct link between the kinematics of drawing actions and the aesthetics of drawing outcomes has not been experimentally demonstrated. Across four experiments, we measured aesthetic responses of students from arts and non-arts backgrounds to drawing movements generated from computational models of human writing. Experiment 1 demonstrated that human-like drawing movements with bell-shaped velocity profiles (Sigma Lognormal [SL] and Minimum Jerk [MJ]) are perceived as more natural and pleasant than movements with a uniform profile, and in both Experiments 1 and 2 movements that were perceived as more natural were also preferred. Experiment 3 showed that this effect persists if lower-level dynamic stimulus features are fully matched across experimental and control conditions. Furthermore, aesthetic preference for human-like movements were associated with greater perceptual fluency in Experiment 3, evidenced by unbiased estimations of the duration of natural movements. In Experiment 4, line drawings with visual features consistent with the dynamics of natural, human-like movements were preferred, but only by art students. Our findings directly link the aesthetics of human action to the visual aesthetics of drawings, but highlight the importance of incorporating artistic expertise into embodied accounts of aesthetic experience.


The clinical and laboratory survey of Iranian patients with hyper-IgE syndrome.
In order to determine the clinical and laboratory findings of Iranian patients with presumed hyper-immunoglobulin E syndrome (HIES), the medical records of 22 patients from 21 unrelated families, who had been registered in the Iranian Primary Immunodeficiency Registry, were observed. The median age of patients at the time of first symptom and at the time of diagnosis was 1 month and 52.5 months, respectively, with a median diagnosis delay of 70 months. 13 families had consanguineous marriages. IgE level was higher than 2000 IU/ml in all patients, ranging from >2000 to 80,000 IU/ml. The most commonly occurring manifestations were: eczema and dermatitis, pneumonia, upper respiratory tract infections, cutaneous abscesses, diarrhoea, deep abscesses, and otitis media. Other less frequent manifestations were: mucocutaneous candidiasis, sinusitis, cutaneous ulcers, Molluscum contagiosum, herpetic keratitis, onychomycosis, conjunctivitis, septic arthritis, and meningitis. Five patients were complicated by bronchiectasis due to recurrent pneumonia and 5 patients died because of severe infections and malignancy. The HIES is a multisystem disorder that affects especially cutaneous, respiratory, skeletal and the immune system. Although HIES is a rare condition, the recurrent infections should always raise a suspicion, which deserves further evaluation for detecting the syndrome.


Pharmacokinetics and Urinary Excretion Mechanism of Orteronel (TAK-700), A Novel 17,20-Lyase Inhibitor, in Animals.
Orteronel is newly identified as a selective 17,20-lyase inhibitor for an agent for castration resistant prostate cancer. The absorption and disposition of [(14)C]orteronel were investigated in rats and monkeys. Orteronel was extensively excreted into rat and monkey urine in an unchanged form after oral administration. The unbound based renal clearances in rats and monkeys were greater than the respective glomerular filtration rates (GFR), suggesting that urinary tubular secretion plays an important role in the renal excretion of orteronel. Therefore, the uptake of [(14)C]orteronel was investigated using rat kidney slices to estimate the contribution of carrier-mediated transport on the urinary tubular secretion. The uptake study using rat kidney slices suggested that the transport of orteronel from the blood circulation to the kidney was mediated by a digoxin sensitive transport system represented by Oatp4c1 and non-saturable components. Furthermore, the saturable component accounted for a limited fraction of the total renal uptake by rat kidney slices. These results suggested that non-saturable uptake mainly contributed to the renal excretion of orteronel in rats.


Minimal effective dose and relapse--double-blind trial: haloperidol decanoate vs. placebo.
Fifty-six chronic schizophrenic patients were randomized into haloperidol decanoate (HD) or placebo groups for 48 weeks of double-blind treatment. The double-blind trial was preceded by a 15-week single-blind run-in period, during which all patients were treated with 60 mg of HD (approximately corresponding to 3.6 mg orally/day) every fourth week (except for the second injection, which was given after 3 weeks). Eight relapses occurred during the run-in period, and seven other patients refused further participation. The remaining 41 patients were then treated double blind, either with HD, 60 mg/4 weeks (18 patients), or with placebo (23 patients). Two patients (11%) in the HD group and 16 (69%) in the placebo group relapsed during the 48-week double-blind period. The plasma concentrations of haloperidol were measured every fourth week in both groups. Steady state was reached after 11 weeks. The mean steady-state level was 6.3 nmol/L. In the placebo group, a 50% decrease in the mean haloperidol plasma concentration was seen 8 weeks after withdrawal of haloperidol. The haloperidol plasma concentration was a predictor of relapse. There was no statistical difference between the treatment groups regarding extrapyramidal symptoms. More biperiden, however, was used in the HD group, while the placebo patients took more sedatives. The results of this study show that the relatively low and fixed dose of 60 mg of HD every fourth week was superior to placebo in preventing relapse in schizophrenia.(ABSTRACT TRUNCATED AT 250 WORDS)


Therapeutic potential of human adipose-derived stem cells (ADSCs) from cancer patients: a pilot study.
Mesenchymal stem cells from adipose tissue (ADSCs) are an important source of cells for regenerative medicine. The therapeutic effect of culture-expanded adipose derived stem cells has been shown; however, optimal xeno-free culture conditions remain to be determined. Cancer patients, specifically those undergoing invasive surgery, constitute a subgroup of patients who could benefit from autologous stem cell transplantation. Although regenerative potential of their ADSCs could be affected by the disease and/or treatment, we are not aware of any study that has evaluated the therapeutic potential of ADSCs isolated from cancer patients in reference to that of ADSCs derived from healthy subjects. Here we report that ADSCs isolated from subabdominal adipose tissue of patients with urological neoplasms yielded similar growth kinetics, presented equivalent mesenchymal surface markers and showed similar differentiation potential into distinct mesodermal cell lineages: adipocytes, chondroblasts and osteoblasts than ADSCs isolated from adipose tissue of age-matched non-oncogenic participants, all under xeno-free growth culture conditions. Molecular karyotyping of patient expanded ADSCs genomes showed no disease-related alterations indicating their safety. In addition, vesicles <100 nm identified as exosomes (EXOs) which may be at least partly responsible for the attributed therapeutic paracrine effects of the ADSCs were effectively isolated from ADSCs and showed equivalent miRNA content regardless they were derived from cancer patients or non-oncogenic participants indicating that the repair capabilities of xeno-free expanded ADSCs are not compromised by patient condition and therefore their xeno-free culture expanded ADSCs should be suitable for autologous stem cell transplantation in a clinical setting.


Self-Enhanced Electrochemiluminescence Imaging System Based on the Accelerated Generation of ROS under Ultrasound.
Electrochemiluminescence (ECL) imaging, as an optical technology, has been developed at full tilt in the field of life science and nanomaterials. However, the relatively low ECL intensity or the high co-reactant concentration needed in the electrochemical reaction blocks its practical application. Here, we developed an ECL imaging system based on the rGO-TiO2-x composite material, where the co-reactant, reactive oxygen species (ROS), is generated in situ under the synergetic effect of of ultrasound (US) and electric irradiation. The rGO-TiO2-x composites facilitate the separation of electron (e-) and hole (h+) pairs and inhibit recombination triggered by external US irradiation due to the high electroconductivity of rGO and oxygen-deficient structures of TiO2, thus significantly boosting ROS generation. Furthermore, the increased defects on rGO accelerate the electron transfer rate, improving the electrocatalytic performance of the composite and forming more ROS. This high ultrasonic-electric synergistic efficacy is demonstrated through the enhancement of photon emission. Compared with the luminescence intensity triggered by US irradiation and electric field, an enhancement of ∼20-fold and 10-fold of the US combined with electric field-triggered emission is observed from this composite. Under the optimized conditions, using dopamine (DA) as a model target, the sensitivity of the US combined ECL strategy for detection of DA is two orders of magnitude higher than that of the ECL method. The successful detection of DA at low concentrations makes us believe that this strategy provides the possibility of applying ECL imaging for cellular single-molecule analysis and cancer therapy.


Structural neuroimaging in adolescents with a first psychotic episode.
The objective of the present study is to replicate findings in first-episode psychosis reporting a smaller volume in brain structures in a population with adolescent onset. Magnetic resonance imaging studies were performed on 23 psychotic adolescents (12-18 years old, 17 males, 6 females) consecutively admitted to an adolescent inpatient unit and on 37 normal controls (13-18 years, 23 males, 14 females) matched for age, sex, and years of education. Diagnosis was made at baseline on the basis of the Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime version and confirmed after 12 months of follow-up. Total brain volume and gray matter, white matter, and cerebrospinal fluid (CSF) volumes of the frontal, parietal, temporal, and occipital lobes were measured bilaterally using a segmentation method based on the Talairach grid system. Male patients showed significantly larger volumes than did male controls in overall CSF and left frontal and right parietal sulci CSF. Male patients also showed significantly lower volumes of gray matter in the right and left frontal lobes. No significant volumetric differences were found in females. There were no differences between individuals with a diagnosis of schizophrenia at follow-up and the rest of the patients. This study suggests that larger CSF and lower gray matter volumes in the frontal lobes may be a nonspecific vulnerability marker for psychosis in male adolescents.


Acute myocardial infarction with no obstructive coronary atherosclerosis: mechanisms and management.
Myocardial infarction (MI) with no obstructive coronary atherosclerosis (MINOCA) is a syndrome with different causes. Its prevalence ranges between 5 and 25% of all MIs. The prognosis is extremely variable, depending on the causes of MINOCA. Clinical history, echocardiography, coronary angiography, and left ventriculography represent the first-level diagnostic investigations. Nevertheless, additional tests are required in order to establish its specific cause, thus allowing an appropriate risk stratification and treatment. We review pathogenesis, diagnosis, prognosis, and therapy of MINOCA and propose an algorithm for its management.


Peptides derived from plasma proteins released by bothropasin, a metalloprotease present in the Bothrops jararaca venom.
Viperid snake venoms contain proteases that affect hemostasis by degrading important proteins such as those that participate in the coagulation cascade. The Bothrops jararaca venom presents as its main components metallo and serine proteases, which comprise around 65% of the venom composition. Bothropasin is a hemorrhagic metalloprotease from the B. jararaca venom which causes disruption of the basement membrane of the vascular endothelium, resulting in bleeding. Although the bothropasin ability to degrade plasmatic and extracellular matrix proteins in vitro has been described, the primary sequence of the released peptides is unknown. This research study presents the peptide identification from both fibrinogen and fibronectin, generated by bothropasin proteolytic activity. Among the fibrinogen derived peptides identified by mass spectrometry, analogous of endogenous products like the fibrinopeptides A and B were found, as well as other sequences described in the literature with vasoactive or antiangiogenic properties. A series of peptides derived from fibronectin by the action of bothropasin were described, and for most of them no biological activity has been described. However, exceptionally a peptide that is known as a bond site for B cells was found. This study indicates that, beyond to the degradation of human proteins, bothropasin can generate bioactive peptides, which may participate in the envenoming process by Bothrops snakes. Also important, the knowledge of the formed peptides, based on the cleavage sites of the hydrolyzed proteins, provided the opportunity to study the primary specificity of bothropasin.


Transient expression and purification of β-caryophyllene synthase in Nicotiana benthamiana to produce β-caryophyllene in vitro.
The sesquiterpene β-caryophyllene is an ubiquitous component in many plants that has commercially been used as an aroma in cosmetics and perfumes. Recent studies have shown its potential use as a therapeutic agent and biofuel. Currently, β-caryophyllene is isolated from large amounts of plant material. Molecular farming based on the Nicotiana benthamiana transient expression system may be used for a more sustainable production of β-caryophyllene. In this study, a full-length cDNA of a new duplicated β-caryophyllene synthase from Artemisia annua (AaCPS1) was isolated and functionally characterized. In order to produce β-caryophyllene in vitro, the AaCPS1 was cloned into a plant viral-based vector pEAQ-HT. Subsequently, the plasmid was transferred into the Agrobacterium and agroinfiltrated into N. benthamiana leaves. The AaCPS1 expression was analyzed by quantitative PCR at different time points after agroinfiltration. The highest level of transcripts was observed at 9 days post infiltration (dpi). The AaCPS1 protein was extracted from the leaves at 9 dpi and purified by cobalt-nitrilotriacetate (Co-NTA) affinity chromatography using histidine tag with a yield of 89 mg kg-1 fresh weight of leaves. The protein expression of AaCPS1 was also confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analyses. AaCPS1 protein uses farnesyl diphosphate (FPP) as a substrate to produce β-caryophyllene. Product identification and determination of the activity of purified AaCPS1 were done by gas chromatography-mass spectrometry (GC-MS). GC-MS results revealed that the AaCPS1 produced maximum 26.5 ± 1 mg of β-caryophyllene per kilogram fresh weight of leaves after assaying with FPP for 6 h. Using AaCPS1 as a proof of concept, we demonstrate that N. benthamiana can be considered as an expression system for production of plant proteins that catalyze the formation of valuable chemicals for industrial applications.


Coordinated regulation of neuronal mRNA steady-state levels through developmentally controlled intron retention.
Differentiated cells acquire unique structural and functional traits through coordinated expression of lineage-specific genes. An extensive battery of genes encoding components of the synaptic transmission machinery and specialized cytoskeletal proteins is activated during neurogenesis, but the underlying regulation is not well understood. Here we show that genes encoding critical presynaptic proteins are transcribed at a detectable level in both neurons and nonneuronal cells. However, in nonneuronal cells, the splicing of 3'-terminal introns within these genes is repressed by the polypyrimidine tract-binding protein (Ptbp1). This inhibits the export of incompletely spliced mRNAs to the cytoplasm and triggers their nuclear degradation. Clearance of these intron-containing transcripts occurs independently of the nonsense-mediated decay (NMD) pathway but requires components of the nuclear RNA surveillance machinery, including the nuclear pore-associated protein Tpr and the exosome complex. When Ptbp1 expression decreases during neuronal differentiation, the regulated introns are spliced out, thus allowing the accumulation of translation-competent mRNAs in the cytoplasm. We propose that this mechanism counters ectopic and precocious expression of functionally linked neuron-specific genes and ensures their coherent activation in the appropriate developmental context.


Millimeter field-of-view miniature two-photon microscopy for brain imaging in freely moving mice.
Development of miniature two-photon microscopy (m2PM) has made it possible to observe fine structure and activity of neurons in the brain of freely moving animals. However, the imaging field-of-view of existing m2PM is still significantly smaller than that of miniature single-photon microscopy. Here we report that, through the design of low-magnification objective, large field-of-view scan lens and small tilt angle microscanner, a 2.5-g m2PM achieved a field-of-view of 1000 × 788 µm2, comparable to that of a typical single-photon miniscope. We demonstrated its capability by imaging neurons, dendrites and spines in the millimeter field-of-view, and simultaneous recording calcium activities, through a gradient-index lens, of approximately 400 neurons in the dorsal hippocampal CA1 in a freely moving mouse. Integrated with a detachable 1.2-g fast z-scanning module, it enables a 1000 × 788 × 500 µm3 volumetric neuronal imaging in the cerebral cortex. Thus, millimeter FOV m2PM provides a powerful tool for deciphering neuronal population dynamics in experimental paradigms allowing for animal's free movement.


[Acute colonic hemorrhage. Importance of a preoperative localization of the origin of bleeding in patients requiring emergency surgery].
Conservative management of distal gastro-intestinal bleeding is successful in most cases; 10% of patients hospitalised with this diagnosis will however undergo emergency surgery. Preoperative localisation of the bleeding site allows to perform a limited, segmental colectomy even in emergency. This has been shown to be associated with a lower operative mortality and morbidity when compared with subtotal colectomy. In this retrospective study we reviewed the notes of 134 patients admitted with lower gastro-intestinal bleeding. 22 of these required more than 4 units of blood transfusion and 12 underwent emergency surgery. Preoperative localisation of the source of bleeding was possible in 7 cases (58%); the remaining 5 underwent a subtotal colectomy. The operative mortality was 8%. The Authors emphasise the importance of an aggressive diagnostic work up in all cases of massive bleeding (i.e. more than 4 units of blood requirement in the first 24 hours following hospitalisation) in order to minimise the number of emergency subtotal colectomy.


Effects of the critical band on auditory-evoked magnetic fields.
Changes in the bandwidth affect the perceived loudness of a stimulus even when the level of the stimulus remains fixed. If the bandwidth of a sound is varied while maintaining the overall intensity, the loudness remains constant as long as the bandwidth is less than the critical bandwidth. If the bandwidth is increased beyond the critical bandwidth, the loudness increases with increasing bandwidth. Human cortical responses as a function of stimulus bandwidth were examined by recording auditory-evoked magnetic fields. The results showed that the N1m magnitudes, that is, the estimated equivalent current dipole moments, increased with increasing bandwidth when the bandwidth was increased beyond the critical bandwidth.


Testing the validity of two putative sympatric species from Sinocyclocheilus (Cypriniformes: Cyprinidae) based on mitochondrial cytochrome b sequences.
There are over 60 species within the freshwater fish genus Sinocyclocheilus (Cypriniformes: Cyprinidae) distributed throughout the Yunnan-Guizhou Plateau and its surrounding areas in China. In recent years, the increasing number of new species described has raised some controversy about the validity of several species within this genus, notably the putative sympatric species pair S. qiubeiensis and S. jiuchengensis. To test the validity of S. qiubeiensis and S. jiuchengensis, we analyzed the complete sequences of the mitochondrial cytochrome b (CYTB) gene of 20 identified species and one outgroup species. Phylogenetic relationships were reconstructed using CYTB with maximum likelihood (ML) and Bayesian inference (BI) methods. Our phylogenetic results showed that all individuals of S. qiubeiensis and S. jiuchengensis clustered in one clade with strong support. In addition, the genetic distance between the two species was 0.11%, within the range observed at the intraspecific level. The most recent common ancestor of S. qiubeiensis and S. jiuchengensis dated back to 0.13 million years ago, indicating little time for complete speciation to have occurred. These results clearly support the viewpoint that S. jiuchengensis is a synonym of S. qiubeiensis.


Additional antacid does not increase the effect of cimetidine in gastric ulcer disease.
Eighty-one adult outpatients with endoscopically confirmed gastric ulceration were treated with cimetidine (1 g/day). In addition, the patients were allocated at random to either intensive antacid treatment or placebo treatment. This part of the study was double-blind. It was found that additional antacid treatment had no effect on ulcer healing and symptoms in cimetidine-treated gastric ulcer patients. The results were similar in patients with corpus ulcers and patients with prepyloric ulcers.


Macular Bruch's membrane defect and dome-shaped macula in high myopia.
To examine an association between macular Bruch's membrane defects (MBMD) and a dome-shaped appearance of the macula (DSM). Retrospective, observational case series study. The study included highly myopic individuals who were consecutively examined between May 2014 and December 2015. The patients underwent swept-source optical coherence tomography (OCT) for visualization of DSM and MBMDs defined as Bruch´s membrane defects located at a distance of maximal 1500 μm from the foveola. Out of 1983 highly myopic eyes (1057 patients), 166 eyes (8.4%; 95% confidence interval (CI):7.2%,9.6%)) showed a DSM and 534 eyes showed a MBMD. In multivariate binary regression analysis, higher prevalence of DSM was associated with a higher prevalence of a MBMD (P<0.001; OR: 1.96; 95%CI: 1.40, 2.75) after adjusting for longer axial length (P<0.001; odds ratio (OR): 1.27; 95%CI: 1.16, 1.38). In eyes with a DSM partially surrounded by a MBMD, the retina, retinal pigment epithelium (RPE) and choroid appeared relatively unchanged in the central region with Bruch´s membrane (BM) preserved. In the ring-like BM-free region surrounding the central prominent island of the DSM, the RPE, the outer and middle retinal layers, the choriocapillaris and the middle-sized choroidal vessel layer were absent. In association with a DSM, three MBMD types were differentiated: MBMDs in patchy chorioretinal atrophy, MBMDs in choroidal neovascularization-related macular atrophy, and MBMDs as temporally extending large parapapillary gamma zone. Presence of a DSM was significantly associated with the presence of MBMDs. The morphology of the DSM in association with MBMDs may be associated with a focal relaxation of the posterior sclera, no longer pushed outward by an expanding BM but allowed to partially bulge inward, leading to the formation of a DSM.


Effects of methyl mercury on murine fetal amino acid uptake, protein synthesis and palate closure.
Methylmercury (MeHg: 5 mg Hg/kg maternal body weight) in 0.13 M NaCl, 0.01 M NaH2PO4-Na2HPO4, pH 7.4 (PBS) administered to gravid CFW mice on day 12, hour 6 (12(6)) of gestation induced a high incidence of cleft palate in fetuses examined on days 15(6) (72%), 16(6) (62%) and 17(6) (40%). Palate closure (100%) in PBS control animals occurred by 14(10). One day post MeHg administration, total fetal protein was decreased 22% while DNA content was unaltered. Protein was maximally decreased (28%) on 14(6) and, thereafter, returned toward control levels. Alterations in DNA content followed a similar pattern; but the maximal decrease (32%) occurred on 15(6). The rate of fetal protein synthesis was depressed 5% at 12(9) and between 20% to 26% from this time to 13(6) (end of observation). The agreement between the calculated decrease in protein synthesis (19%) and the measured decrease in protein content (22%) suggests that a reduction in protein synthesis is responsible for the decreased fetal protein content. Placental blood flow and fetal water space, measured with 3H--H2O at 12(18), were not affected by MeHg treatment. However, fetal free amino acid concentrations at 12(18) were generally decreased (alanine, 23.0%; valine, 9.7%; methionine, 22.6%; isoleucine, 12.0%; leucine, 18.2%) while uptake of the non-metabolizable amino acid, 14C-cycloleucine, was decreased 23%. From this, it is concluded that the growth inhibitory effects of MeHg are related, at least in part, to impaired placental/fetal transfer of amino acids.


Modification of oligosaccharide antenna flexibility induced by exoglycosidase trimming.
We have investigated the solution conformation of a triantennary glycopeptide using resonance energy transfer [Rice et al. (1991) Biochemistry 30, 6646-6655]. Triantennary glycopeptide was derivatized with a donor fluorophore on the N-terminus and with an acceptor fluorophore attached individually to each terminal galactose residue, resulting in three isomeric donor-acceptor pairs. Time-resolved energy-transfer experiments revealed two distinct donor-acceptor distance populations for two of the three antennae of the oligosaccharide. An extended conformation and a folded conformation were detected for the two flexible antennae whereas the third antenna on the oligosaccharide was rigid, containing only an extended conformer. The ratios of the extended to folded conformers of the two flexible antennae were reversibly modulated by temperature, allowing the calculation of delta H and delta S for the conformational change [Wu et al. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 9355-9359]. In the present study, we have trimmed the fluorescent glycopeptides with exoglycosidases which specifically removed the unmodified antennae of the oligosaccharide. The resulting single-chain isomeric glycopeptides each contained identical core sugar residues and a terminally located donor and acceptor, but differed only in the linkage configuration of the sugar residues. Analysis of these glycopeptides by time-resolved energy transfer indicated that each antenna of the oligosaccharide is, by itself, maintained exclusively in the extended conformation. Temperature modulation failed to induce antenna folding as was previously observed for the complete triantennary structure. These data suggest that interantenna interactions modulate the conformation of individual antenna of complex oligosaccharides.


[Role of macrophages and fibronectin in vitreoretinal proliferation].
Proliferative vitreoretinopathy (PVR) is characterized by the periretinal proliferation of nonneoplastic cells complicated by traction retinal detachment. In this study, we report the demonstration of macrophages, which are thought to be involved in the etiology of this intraocular disease, in human periretinal membranes by an immunochemical staining procedure using a monoclonal anti-human macrophage antibody. Fibronectin, a high-molecular protein of plasma and extracellular matrix, may be visualized accordingly and could act concomitantly with macrophages as an important factor in PVR. Fibronectin is shown to be present in high quantities (Elisa) in vitreous aspirates from patients with PVR. We hypothesize that the interaction of macrophages and fibronectin may be a crucial step in the development of PVR.


Characterization of a chromosomal nickel resistance determinant from Klebsiella oxytoca CCUG 15788.
Klebsiella oxytoca CCUG 15788 is resistant to Ni2+ at a concentration of 10 mM and grows in an inducible manner when exposed to lower concentrations of Ni2+. The complete genomic sequence of a 4.2-kb HindIII-digested fragment of this strain was determined from genomic DNA. It was shown to contain four nickel resistance genes (nirA, nirB, nirC, and nirD) encoding transporter and transmembrane proteins for nickel resistance. When the plasmid pKOHI4, encoding nirABCD, was transformed into Escherichia coli JM109, the cells were able to grow in Tris-buffered mineral medium containing 3 mM nickel. TnphoA'-1 insertion mutants in the four nickel genes nirA, nirB, nirC, and nirD showed nickel sensitivity. The nir genes were heterogeneously expressed in E. coli, suggesting functional roles of these genes in nickel resistance.


Association of Tumour Microenvironment with Protein Glycooxidation, DNA Damage, and Nitrosative Stress in Colorectal Cancer.
In this study, we evaluated the total antioxidant capacity, nitrosative stress, and protein/DNA oxidation and glycoxidation products in patients with colorectal cancer regarding histopathological parameters associated with the tumour microenvironment, such as inflammatory infiltration and tumour budding and compare all determined parameters between tumours located in the right and left side of the colon and normal mucosa. Ferric reducing antioxidant power (FRAP), nitrosative stress (myeloperoxidase (MPO), nitrogen oxide (NO), peroxynitrite, and nitrotyrosine), protein oxidation products (protein carbonyls (PC), total thiols, and ischemia modified albumin (IMA)), protein glycooxidation products (tryptophan, kynurenine, N-formylkynurenine, dityrosine, Amadori product, advanced glycation end products (AGE)) and 8-hydroxydeoxyguanosine (8-OHdG) were measured in homogenates from normal and cancerous tissue of 30 patients with colorectal cancer. Levels of FRAP (p=0.0009), IMA (p=0.0002), kynurenine (p<0.0001), N-formylkynurenine (p<0.0001), dityrosine (p<0.0001), Amadori products (p=0.0024), AGE (p<0.0001), MPO (p<0.0001), NO (p<0.0001) and nitrotyrosine (p=0.0011) were increased, whereas PC (p=0.0004), tryptophan (p<0.0001), 8-OHdG (p<0.0001) and peroxynitrite (p=0.0003) were decreased in the left-side tumour compared to the right-side tumour and normal mucosa. Our results showed that colorectal cancer is related with disturbances in antioxidant defense and increased oxidative and nitrosative damages to proteins and DNA. These parameters may be useful for evaluation the progression and differentiation of the tumour location. We also demonstrated that redox indicators may depend on the histological type of the tumour and may influence tumour invasion depth, presence of lymph node and distant metastasis, vascular and neural invasion, inflammatory infiltration, and tumour budding, which are part of the tumour microenvironment.


[Preoperative radiotherapy of brain tumors].
Irradiation prior to radical surgical removal has been performed 51 times in 48 cases of brain tumor over the last 7 years. Through this procedure, all cases have attained good results in both clinical and CT evaluations. In the course of radiotherapy, various doses of corticosteroids and/or ventricular drainage were applied to prevent dangerous brain swelling and hydrocephalus. Total radiation doses were 40-50 Gy in primary cases and 30-40 Gy in recurrent cases. Through this preoperative radiotherapy, tumors showed reduction of their feeding vessels (78.9%) on angiography and/or became necrotized with cyst formation. This facilitated case of surgery and permitted radical resection of these tumors (41%). Especially, in cases of germ cell tumors of mixed type and medulloblastomas, this procedure permitted radical resection in 75% and 100% of cases, respectively. Among glioblastomas and grade 3 astrocytomas, 21.7% were radically resected and the patients involved showed a good relative survival rate. This method may actually facilitate radical cure of malignant brain tumors soon.


Intra-Lot and Inter-Lot Variability in Ceftaroline Fosamil.
Intra-lot and inter-lot variability in ceftaroline fosamil was detected in the DQS using Fourier transform near-infrared spectrometry. One vial of 18 sampled from Lot0013E06 appeared 9.0 SDs from the center of the rest of the vials. Six lots in the spectral library clustered in two large groups.


Induced Pluripotent Stem Cells: Reprogramming Platforms and Applications in Cell Replacement Therapy.
The generation of induced pluripotent stem cells (iPSCs) from differentiated mature cells is one of the most promising technologies in the field of regenerative medicine. The ability to generate patient-specific iPSCs offers an invaluable reservoir of pluripotent cells, which could be genetically engineered and differentiated into target cells to treat various genetic and degenerative diseases once transplanted, hence counteracting the risk of graft versus host disease. In this context, we review the scientific research streams that lead to the emergence of iPSCs, the roles of reprogramming factors in reprogramming to pluripotency, and the reprogramming strategies. As iPSCs serve tremendous correction potentials for various diseases, we highlight the successes and challenges of iPSCs in cell replacement therapy and the synergy of iPSCs and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing tools in therapeutics research.


Copper(II).bleomycin, iron(III).bleomycin, and copper(II).phleomycin: comparative study of deoxyribonucleic acid binding.
The kinetics and mechanism of binding of Cu-(II).bleomycin, Fe(III).bleomycin, and Cu(II).phleomycin to DNA were studied by using fluorometry, equilibrium dialysis, electric dichroism, and temperature-jump and stopped-flow spectrophotometry. The affinity of Cu(II).bleomycin for DNA was greater than that of metal-free bleomycin but less than that of Fe(III).bleomycin. Cu(II).bleomycin exhibited a two-step binding process, with the slow step indicating a lifetime of 0.1 s for the Cu(II).bleomycin.DNA complex. Fe(III).bleomycin binding kinetics indicated the presence of complexes having lifetimes of up to 22 s. DNA was lengthened by 4.6 A/molecule of bound Cu(II).bleomycin and by 3.2 A/bound Fe(III).bleomycin but not at all by Cu(II).phleomycin, suggesting that both bleomycin complexes intercalate while the phleomycin complex does not. However, phleomycin exhibited nearly the same specificity of DNA base release as bleomycin. These results suggest that the coordinated metal ion plays a major role in the binding of metal-bleomycin complexes to DNA but that intercalation is neither essential for DNA binding and degradation nor primarily responsible for the specificity of DNA base release by these drugs.


Reactive oxygen species as therapeutic targets in pulmonary hypertension.
Pulmonary hypertension (PH) is characterized by a progressive elevation of pulmonary arterial pressure due to alterations of both pulmonary vascular structure and function. This disease is rare but life-threatening, leading to the development of right heart failure. Current PH treatments, designed to target altered pulmonary vascular reactivity, include vasodilating prostanoids, phosphodiesterase-5 inhibitors and endothelin-1 receptor antagonists. Although managing to slow the progression of the disease, these molecules still do not cure PH. More effective treatments need to be developed, and novel therapeutic strategies, targeting in particular vascular remodelling, are currently under investigation. Reactive oxygen species (ROS) are important physiological messengers in vascular cells. In addition to atherosclerosis and other systemic vascular diseases, emerging evidence also support a role of ROS in PH pathogenesis. ROS production is increased in animal models of PH, associated with NADPH oxidases increased expression, in particular of several Nox enzymes thought to be the major source of ROS in the pulmonary vasculature. These increases have also been observed in vitro and in vivo in humans. Moreover, several studies have shown either the deleterious effect of agents promoting ROS generation on pulmonary vasculature or, conversely, the beneficial effect of antioxidant agents in animal models of PH. In these studies, ROS production has been directly linked to pulmonary vascular remodelling, endothelial dysfunction, altered vasoconstrictive responses, inflammation and modifications of the extracellular matrix, all important features of PH pathophysiology. Altogether, these findings indicate that ROS are interesting therapeutic targets in PH. Blockade of ROS-dependent signalling pathways, or disruption of sources of ROS in the pulmonary vasculature, targeting in particular Nox enzymes, represent promising new therapeutic strategies in this disease.


Physical inactivity and low fitness deserve more attention to alter cancer risk and prognosis.
Sedentary lifestyle is associated with elevated cancer risk whereas regular physical activity (PA) and high cardiorespiratory fitness (CRF) have the opposite effect, with several biologic mechanisms mediating such associations. There is a need for lifestyle interventions aimed at increasing the PA levels and CRF of the general population and particularly cancer survivors. Furthermore, provocative data suggest a dose-dependent benefit of increasing levels of PA and/or CRF against cancer risk or mortality. Thus, current PA guidelines (≥150 min/wk of moderate-to-vigorous PA) may not be sufficiently rigorous for preventing cancer nor for extending cancer survivorship. Research targeting this issue is urgently needed. Promoting regular PA along with monitoring indicators of CRF and adiposity may provide powerful strategies to prevent cancer in populations, help patients with cancer more effectively deal with their disease and enhance secondary prevention programs in those who are affected by cancer.


Topical 5% tranexamic acid with 30% glycolic acid peel: An useful combination for accelerating the improvement in melasma.
Recently, topical Tranexamic acid (TXA) has been used in melasma management. On detail search of literature, this may be the first study-assessing efficacy on combining of Topical TXA with GA peel in melasma. The aim of this study is to assess efficacy, safety, and improvement in quality of life index on combining 30% GA peel with 5% TXA solution topically in melasma of epidermal type. Sixty patients of epidermal melasma were included in the study and were categorized into two groups: Combination group was treated with 30% GA peel at 2 weekly intervals with 5% TXA solution applied twice daily and Control group was treated with only 30% GA peel every 2 weeks for 12 weeks. Melasma area severity index (MASI) was used for assessing clinical improvement. Hi-MELASQOL and HRQOL scales were used to measure Melasma related quality of life and were compared between both groups. At each visit, adverse effects were noted. A significantly decreasing trend was seen regarding the MASI score when compared within the group, but the difference was statistically not significant between the two groups at 12 weeks. However, significant reduction in MASI score was attained earlier in the combination group than the control group. Similarly, there was significant improvement in Hi-MELASQOL and HRQOL in both the groups, but the difference between them was statistically not significant. Side effects experienced by patients in both groups were trivial and did not require stoppage of therapy. This study concluded that topical TXA with GA peel has comparable result with GA peel alone, but the therapeutic response was achieved in patients of combination group earlier in comparison to control group patients.


The electronic spectrum of the C2P free radical and a Renner-Teller analysis of the 2Delta and X 2Pi electronic states.
Subsequent to our spectroscopic detection of the C(2)X(X=P,As) free radicals [F. X. Sunahori et al., J. Am. Chem. Soc. 129, 9600 (2007)], we have studied the electronic spectrum of the (2)Delta(i)-X (2)Pi(r) system of the jet-cooled C(2)P free radical in the 490-630 nm region. The high-resolution laser-induced fluorescence spectrum of the two spin components of the 0(0) (0) band of (12)C(2)P has been recorded, and the rotational and spin-orbit coupling constants have been determined for both electronic states. The Renner-Teller effect has been observed in both the (2)Pi and the (2)Delta states, and the vibrational structure has been assigned. For the ground state, all of the observed levels up to 3500 cm(-1) were fitted with a standard Renner-Teller model. The excited (2)Delta state vibrational levels were successfully fitted using literature energy level expressions derived from perturbation theory, yielding vibrational and Renner-Teller parameters for both (12)C(2)P and (13)C(2)P. The molecular structure of C(2)P in the ground and excited states has also been estimated and compared to ab initio calculations and the geometries of similar molecules.


Erratum: Obtaining High-Quality Transcriptome Data from Cereal Seeds by a Modified Method for Gene Expression Profiling.
An erratum was issued for: Obtaining High-Quality Transcriptome Data from Cereal Seeds by a Modified Method for Gene Expression Profiling. Two author names were updated. The names were corrected from: Christianne Seiler Nese Sreeenivasulu to: Christiane Seiler Nese Sreenivasulu.


Fengreqing Oral Liquid Exerts Anti-Inflammatory Effects by Promoting Apoptosis and Inhibiting PI3K/AKT and NF-κB Signaling Pathways.
Fengreqing oral liquid (FOL), a Chinese patent drug frequently used in clinical practice in China, is effective in treating inflammatory diseases of the upper respiratory tract such as colds and flu. However, its anti-inflammatory effects and mechanisms remain to be elucidated. In this study, the anti-inflammatory effects of FOL and its mechanisms on PI3K/AKT and NF-κB signaling pathways in LPS-induced RAW264.7 cells were explored, as well as the regulatory effect of FOL on apoptosis. In addition, the potential of FOL for the treatment of acute lung injury was explored in LPS-induced ALI mice. The results showed that treatment with FOL significantly reduced the levels of interleukin 1β (IL-1β), interleukin 6 (IL-6), nitric oxide (NO), and tumor necrosis factor α (TNF-α) in the supernatant of LPS-induced RAW264.7 cells, and also significantly reduced the phosphorylated protein levels of PI3K and AKT in the PI3K/AKT signaling pathway and also protein levels of NF-κB p50, phosphorylated NF-κB p65, and IκBα in the NF-κB signaling pathway. In addition, the results showed that FOL induced apoptosis in LPS-induced RAW264.7 cells at the level of 80%-90%, and significantly increased the protein expression levels of the pro-apoptotic Bax and cleaved-caspase-3. In LPS-induced ALI mice, FOL administration showed inhibition of IL-1β, IL-6, and TNF-α in Bronchoalveolar lavage fluid (BALF) and decreased protein expression levels of PI3K, AKT, NF-κB p50, and NF-κB p65, and elevated protein expression levels of Bax and cleaved-caspase-3 significantly. These results suggest that FOL may exert anti-inflammatory effects by inhibiting the PI3K/AKT signaling pathway to promote apoptosis and leading to attenuated activation of the NF-κB signaling pathway.


Transesophageal echocardiographic diagnosis of left atrial mass as thrombus by demonstrating its attachment to the patch closing atrial septal defect: a case report.
A 65-year-old-man was referred for echocardiographic examination because of palpitations. He had a history of an atrial septal defect surgically treated with an artificial synthetic polyester textile fiber patch. TTE showed a large, mobile mass in a dilated left atrium. The attachment site of the mass was not clear by transthoracic approach, and it was difficult to diagnose the mass as a thrombus or a myxoma from the nature of the echocardiographic findings. Transesophageal echocardiography clearly demonstrated that the mass was attached to the patch closing the atrial septal defect and the mass was confirmed as thrombus at surgery. A patient with left atrial thrombus in whom transesophageal echocardiographic demonstration of the attachment of the left atrial mass to a patch closing an atrial septal defect served as an essential clue leading to accurate diagnosis is reported. Transesophageal echocardiography is feasible to evaluate the attachment site of a left atrial mass and to lead to an accurate diagnosis.


CLIC1 Expression in Skin Biopsies from Patients With Rheumatoid and Psoriatic Arthritis as a Potential Tool to Predict Therapy Response.
Chloride intracellular channel protein 1 (CLIC1) activates inflammasomes in rheumatoid (RA) and psoriatic (PsA) arthritis. We studied CLIC1 expression in RA and PsA patients' skin with vasculitis and its variability depending on the therapy used. CLIC1 immunoexpression was evaluated in the vascular (CLIC1-V) and stromal (CLIC1-S) compartments of the RA and PsA skin biopsies of patients treated with methotrexate (MTX), leflunomid (LFN), corticotherapy (CT), or biological therapies. MTX significantly reduced CLIC1-S expression (p=0.016), whereas LFN decreased CLIC1-V (p<0.001). LFN therapy duration also correlated with CLIC1-V (p<0.001). CT decreased CLIC1-S expression (p=0.006). CLIC1-S expression persisted in skin biopsies despite of erythrocyte sedimentation rate (ESR, p=0.018) and C reactive protein (CRP, p=0.0026) normalisation. For PsA, CLIC1-S expression significantly related to MTX (p<0.022). Both CLIC1-S (p<0.001) and CLIC1-V (p=0.007) decreased by biological therapies in RA. CLIC1 expression is strongly influenced by the therapy used. Our data strongly support the extensive evaluation of CLIC1 in RA as a potential marker of inflammation and tool to predict therapy response.


Phototransformative Supramolecular Assembly of Amphiphilic Diarylethenes Realized by a Combination of Photochromism and Lower Critical Solution Temperature Behavior.
Amphiphilic diarylethenes bearing octyloxycarbonyl and N-octylcarbamoyl groups have been designed and synthesized. These ester- and amide-linked compounds form micrometer-sized supramolecular assemblies in water, and these assemblies exhibit photoinduced macroscopic morphological transformations upon alternate irradiation with UV and visible light. The ester-linked diarylethene showed a transformation between colorless spheres and a red-purple hazy fringe, whereas the microspheres of the amide-linked diarylethene showed changes in color, size, and shape, but the spheres did not show division. TEM images revealed that the spheres of the open-ring isomers have coacervate structures, with bicontinuous aqueous and organic phases. The closed-ring isomers of the ester- and amide-linked compounds were found to form nanofibers and thin layers, respectively. These compounds showed absorption spectral shifts at temperatures corresponding to the lower critical solution temperature (LCST) transition. This morphological transformation can be rationalized as the photoinduced phase transition between the high- and low-temperature phases of the LCST transition. These results open up a new avenue for the design of phototransformative supramolecular assemblies based on a combination of photochromism and LCST behavior.


Pyridine-Spiropyran Derivative as a Persistent, Reversible Photoacid in Water.
A highly versatile water-soluble pyridine-spiropyran photoswitch is reported which functions as photoacid in a wide pH range. Under neutral conditions, the open-ring merocyanine (MC-) exists to 48% and closes quantitatively by irradiation with visible light, while the reverse reaction occurs rapidly in the dark or by irradiation at 340 nm. The different pKa of the pyridine nitrogen in the closed spiropyran (4.8) and open merocyanine form (6.8) leads to a reversible proton release in a pH range of 3-7. Only negligible hydrolytic decomposition was observed in the pH range from 1 to 12. The application of potentially harmful UV light can be circumvented due to the fast thermal ring-opening except for pH values below 3. Its photoacidic properties make this compound an effective pH-regulating photoswitch in water and enable controlled proton-transfer processes for diverse applications. Additionally, all of the involved protonated states of the compound exhibit discriminative fluorescence features within certain pH ranges, which even expands its utility to a light-controllable, pH-sensitive fluorophore.


Use of toxicogenomics for identifying genetic markers of pulmonary oedema.
This study was undertaken primarily to identify genetic markers of oedema and inflammation. Mild pulmonary injury was induced following the instillation of the oedema-producing agent, bleomycin (0.5 units). Oedema was then confirmed by conventional toxicology (lavage protein levels, free cell counts and lung/body weight ratios) and histology 3 days post-bleomycin instillation. The expression profile of 1176 mRNA species was determined for bleomycin-exposed lung (Clontech Atlas macroarray, n=9). To obtain pertinent results from these data, it was necessary to develop a simple, effective method for bioinformatic analysis of altered gene expression. Data were log10 transformed followed by global normalisation. Differential gene expression was accepted if: (a) genes were statistically significant (P < or = 0.05) from a two-tailed t test; (b) genes were consistently outside a two standard deviation (SD) range from control levels. A combination of these techniques identified 31 mRNA transcripts (approximately 3%) which were significantly altered in bleomycin treated tissue. Of these genes, 26 were down-regulated whilst only five were up-regulated. Two distinct clusters were identified, with 17 genes classified as encoding hormone receptors, and nine as encoding ion channels. Both these clusters were consistently down-regulated. The magnitude of the changes in gene expression were quantified and confirmed by Q-PCR (n = 6), validating the macroarray data and the bioinformatic analysis employed. In conclusion, this study has developed a suitable macroarray analysis procedure and provides the basis for a better understanding of the gene expression changes occurring during the early phase of drug-induced pulmonary oedema. This work has been presented orally, in part at the British Association for Lung Research Summer Meeting, University of Brighton, 3-5 September, 2003 and in full at the British Toxicology Society Annual Congress, Heriot Watt University, Edinburgh, 21-24 April 2004.


IFN-beta 2/IL-6 augments the activity of human natural killer cells.
MHC nonrestricted cytotoxic cells play an important role in the killing of tumor cells in vitro and potentially in vivo. The activity of these cells is regulated by several cytokines such as IL-2 and IFN. In the present study we provide first evidence that IL-6 significantly augments the cytotoxic activity of human NK cells. IL-6 is produced by many different cells and is also known as IFN-beta 2, B cell stimulatory factor 2, hybridoma growth factor, hepatocyte-stimulating factor, and 26 kDa protein. IL-6 stimulates the activity of human CD3- NK cells but not that of CD3+ non-MHC-restricted cytotoxic T lymphocytes. As is the case with IL-2, the IL-6-mediated augmented cytotoxicity was a result of a more efficient lysis, but was not caused by an increased effector to target cell binding. Moreover, the effect of IL-6 on NK cell activity was blocked by a mAb directed against IL-2, and IL-6 itself was found to be a potent inducer of IL-2 production in cultured human PBMC. Thus it may be concluded that IL-6 enhances the cytotoxic activity of NK cells via IL-2. This newly recognized property of IL-6, which is produced by almost any cell, may be of importance in host defense against microbes and malignancies and therefore could contribute to improve the adoptive immunotherapy by using lymphokine-activated killer cells.


Mixture toxicity in the marine environment: Model development and evidence for synergism at environmental concentrations.
Little is known about the effect of metal mixtures on marine organisms, especially after exposure to environmentally realistic concentrations. This information is, however, required to evaluate the need to include mixtures in future environmental risk assessment procedures. We assessed the effect of copper (Cu)-Nickel (Ni) binary mixtures on Mytilus edulis larval development using a full factorial design that included environmentally relevant metal concentrations and ratios. The reproducibility of the results was assessed by repeating this experiment 5 times. The observed mixture effects were compared with the effects predicted with the concentration addition model. Deviations from the concentration addition model were estimated using a Markov chain Monte-Carlo algorithm. This enabled the accurate estimation of the deviations and their uncertainty. The results demonstrated reproducibly that the type of interaction-synergism or antagonism-mainly depended on the Ni concentration. Antagonism was observed at high Ni concentrations, whereas synergism occurred at Ni concentrations as low as 4.9 μg Ni/L. This low (and realistic) Ni concentration was 1% of the median effective concentration (EC50) of Ni or 57% of the Ni predicted-no-effect concentration (PNEC) in the European Union environmental risk assessment. It is concluded that results from mixture studies should not be extrapolated to concentrations or ratios other than those investigated and that significant mixture interactions can occur at environmentally realistic concentrations. This should be accounted for in (marine) environmental risk assessment of metals. Environ Toxicol Chem 2017;36:3471-3479. © 2017 SETAC.


Tethering human immunodeficiency virus 1 integrase to a DNA site directs integration to nearby sequences.
Certain retrovirus and retrotransposons display strong biases in the selection of host DNA sites for integration. To probe the possibility that simple tethering of the retroelement integrase protein to a target DNA site is sufficient to direct integration, the activities of a hybrid composed of human immunodeficiency virus 1 integrase and lambda repressor were analyzed. In in vitro reactions containing several target DNAs, the lambda repressor-integrase hybrid was found to direct integration selectively to targets containing lambda operators. Addition of lambda repressor blocked selective integration, indicating that binding to the operators was required. The lambda repressor-integrase hybrid protein directed integration primarily to sites near the operators on the same face of the B-DNA helix, indicating that target DNA was probably captured by looping out the intervening sequences. Such hybrid integrase proteins may be useful for directing retroviral integration to specific sequences in vivo.


Inactivation of interleukin-30 in colon cancer stem cells via CRISPR/Cas9 genome editing inhibits their oncogenicity and improves host survival.
Progression of colorectal cancer (CRC), a leading cause of cancer-related death worldwide, is driven by colorectal cancer stem cells (CR-CSCs), which are regulated by endogenous and microenvironmental signals. Interleukin (IL)-30 has proven to be crucial for CSC viability and tumor progression. Whether it is involved in CRC tumorigenesis and impacts clinical behavior is unknown. IL30 production and functions, in stem and non-stem CRC cells, were determined by western blot, immunoelectron microscopy, flow cytometry, cell viability and sphere formation assays. CRISPR/Cas9-mediated deletion of the IL30 gene, RNA-Seq and implantation of IL30 gene transfected or deleted CR-CSCs in NSG mice allowed to investigate IL30's role in CRC oncogenesis. Bioinformatics and immunopathology of CRC samples highlighted the clinical implications. We demonstrated that both CR-CSCs and CRC cells express membrane-anchored IL30 that regulates their self-renewal, via WNT5A and RAB33A, and/or proliferation and migration, primarily by upregulating CXCR4 via STAT3, which are suppressed by IL30 gene deletion, along with WNT and RAS pathways. Deletion of IL30 gene downregulates the expression of proteases, such as MMP2 and MMP13, chemokine receptors, mostly CCR7, CCR3 and CXCR4, and growth and inflammatory mediators, including ANGPT2, CXCL10, EPO, IGF1 and EGF. These factors contribute to IL30-driven CR-CSC and CRC cell expansion, which is abrogated by their selective blockade. IL30 gene deleted CR-CSCs displayed reduced tumorigenicity and gave rise to slow-growing and low metastatic tumors in 80% of mice, which survived much longer than controls. Bioinformatics and CIBERSORTx of the 'Colorectal Adenocarcinoma TCGA Nature 2012' collection, and morphometric assessment of IL30 expression in clinical CRC samples revealed that the lack of IL30 in CRC and infiltrating leucocytes correlates with prolonged overall survival. IL30 is a new CRC driver, since its inactivation, which disables oncogenic pathways and multiple autocrine loops, inhibits CR-CSC tumorigenicity and metastatic ability. The development of CRISPR/Cas9-mediated targeting of IL30 could improve the current therapeutic landscape of CRC.


Toward Spatially Regulated Division of Protocells: Insights into the E. coli Min System from in Vitro Studies.
For reconstruction of controlled cell division in a minimal cell model, or protocell, a positioning mechanism that spatially regulates division is indispensable. In Escherichia coli, the Min proteins oscillate from pole to pole to determine the division site by inhibition of the primary divisome protein FtsZ anywhere but in the cell middle. Remarkably, when reconstituted under defined conditions in vitro, the Min proteins self-organize into spatiotemporal patterns in the presence of a lipid membrane and ATP. We review recent progress made in studying the Min system in vitro, particularly focusing on the effects of various physicochemical parameters and boundary conditions on pattern formation. Furthermore, we discuss implications and challenges for utilizing the Min system for division site placement in protocells.


Interventions Targeting Health and Wellbeing among Nail Salon Workers: A Scoping Review.
To identify existing interventions targeting the health and wellbeing of nail salon workers. Arksey and O'Malley's framework on reviews guided this project. The databases MEDLINE, EMBASE, Scopus, CINAHL, and Web of Science were searched. A Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) chart was used to capture the inclusion protocol. Four interventions were identified, of which one was a randomized controlled trial. Across the four interventions, there were about 424 participants that were recruited through cosmetology schools and 34 from nail salons. Interventions varied from pamphlet distribution and inspections to full-day training sessions. Intervention effectiveness was determined via pre- and post-tests and infractions frequency. Maximum follow-up time was 8 months. Outcomes included increased knowledge on exposures, fewer infractions, increased adherence to infection control practices and use of personal protective equipment, and behavior and attitude changes. There is a need for further research on interventions targeting vulnerable workers in nail salons who face numerous hazards on the job. Two of the four interventions targeted nail technician trainees and their instructors, indicating the potential of early interventions before the workers officially enter the workforce.


A phase II study of antibody-drug conjugate, TAK-264 (MLN0264) in previously treated patients with advanced or metastatic pancreatic adenocarcinoma expressing guanylyl cyclase C.
Background This phase II open-label, multicenter study evaluated the efficacy, safety, and tolerability of TAK-264 in previously treated patients with advanced or metastatic pancreatic adenocarcinoma expressing guanylyl cyclase C (GCC). Methods Patients with advanced or metastatic pancreatic adenocarcinoma expressing GCC (H-score ≥ 10) received TAK-264 1.8 mg/kg on day 1 of a 21-day cycle as a 30-min intravenous infusion for up to 1 year or until disease progression or unacceptable toxicity. The primary objective was overall response rate (ORR [complete response + partial response (PR)]). Secondary objectives included evaluations of the safety and pharmacokinetic profile of TAK-264 (NCT02202785). Results 43 patients were enrolled and treated with 1.8 mg/kg TAK-264: 11, 15, and 17 patients with low, intermediate, and high GCC expression, respectively. Median number of treatment cycles received was two (range 1-10). The ORR was 3%, including one patient with intermediate GCC expression who achieved a PR. All patients experienced ≥1 adverse events (AE). The majority of patients experienced grade 1/2 AEs affecting the gastrointestinal tract. Fifteen (35%) patients experienced ≥grade 3 drug-related AEs; five (12%) patients had a serious AE. The most common (≥10% of patients) all-grade drug-related AEs were nausea (33%), fatigue (28%), neutropenia (23%), decreased appetite (23%), vomiting (16%), asthenia (16%), and alopecia (14%). Conclusions TAK-264 demonstrated a manageable safety profile; however, the low efficacy of TAK-264 observed in this study did not support further clinical investigation.


Genes and genomes and unnecessary complexity in precision medicine.
The sequencing of the human genome heralded the new age of 'genetic medicine' and raised the hope of precision medicine facilitating prolonged and healthy lives. Recent studies have dampened this expectation, as the relationships among mutations (termed 'risk factors'), biological processes, and diseases have emerged to be more complex than initially anticipated. In this review, we elaborate upon the nature of the relationship between genotype and phenotype, between chance-laden molecular complexity and the evolution of complex traits, and the relevance of this relationship to precision medicine. Molecular contingency, i.e., chance-driven molecular changes, in conjunction with the blind nature of evolutionary processes, creates genetic redundancy or multiple molecular pathways to the same phenotype; as time goes on, these pathways become more complex, interconnected, and hierarchically integrated. Based on the proposition that gene-gene interactions provide the major source of variation for evolutionary change, we present a theory of molecular complexity and posit that it consists of two parts, necessary and unnecessary complexity, both of which are inseparable and increase over time. We argue that, unlike necessary complexity, comprising all aspects of the organism's genetic program, unnecessary complexity is evolutionary baggage: the result of molecular constraints, historical circumstances, and the blind nature of evolutionary forces. In the short term, unnecessary complexity can give rise to similar risk factors with different genetic backgrounds; in the long term, genes become functionally interconnected and integrated, directly or indirectly, affecting multiple traits simultaneously. We reason that in addition to personal genomics and precision medicine, unnecessary complexity has consequences in evolutionary biology.


Antisense-mediated melanoma inhibitor of apoptosis protein downregulation sensitizes G361 melanoma cells to cisplatin.
Malignant melanoma is an aggressive form of skin cancer that is highly resistant to conventional therapies. The melanoma inhibitor of apoptosis protein is a potent inhibitor of apoptosis and is overexpressed in melanoma cells, but undetectable in most normal tissues including melanocytes. We designed 20-mer phosphorothioate antisense oligonucleotides complementary to five putatively single-stranded sites on the melanoma inhibitor of apoptosis protein mRNA and investigated their ability to sensitize G361 melanoma cells to cisplatin. Inhibition of melanoma inhibitor of apoptosis protein mRNA and protein expression were measured by real-time polymerase chain reaction and immunoblotting. Cell viability and apoptosis were quantitated by colorimetric viability assays and by annexin V staining, respectively. Oligonucleotide M706 was identified as the most efficient antisense sequence which downregulated melanoma inhibitor of apoptosis protein mRNA and protein levels in G361 cells by 68 and 78%, respectively. The specificity of target downregulation was confirmed using scrambled sequence control oligonucleotides that only marginally decreased melanoma inhibitor of apoptosis protein expression. Whereas downregulation of melanoma inhibitor of apoptosis protein moderately inhibited cell growth by 26%, in combination with cisplatin, this resulted in a supra-additive effect with almost 57% reduction in G361 cell viability compared with cisplatin alone (17%) (P<0.05). Cell death was mainly due to apoptosis as demonstrated by a 3- to 4-fold increase in annexin V-positive cells and typical morphological changes compared with controls. In summary, we describe a new antisense oligonucleotide that efficiently downregulates melanoma inhibitor of apoptosis protein expression and sensitizes melanoma cells to cisplatin.


Tetracyclines inhibit human synovial collagenase in vivo and in vitro.
To determine if tetracyclines can inhibit human synovial collagenase from rheumatoid tissue, paired synovial tissue (or synovial fluid) was collected from 7 patients before and after oral administration of minocycline (100 mg BID) for 10 days. With each patient serving as his own control, the postminocycline collagenase activities fell an average of 67% from pretreatment values. Qualitative SDS-PAGE revealed decreased loss of alpha collagen components and reduced formation of alpha A digestion fragments. Addition of minocycline or a chemically modified tetracycline to synovial culture media in vitro profoundly inhibited collagenase activity. Further study of this action of tetracyclines could serve as a probe of the role of collagenase in rheumatoid arthritis and lead to development of agents capable of modifying the tissue destructive actions of collagenase.


Retrospective analysis of the endometrial preparation protocols for frozen-thawed embryo transfer cycles in women with endometriosis.
There was inconsistency in optimal endometrial preparation protocol for frozen-thawed embryo transfer (FET) in patients with endometriosis. We conducted this study to investigate the effect of different endometrial preparation protocols on the pregnancy outcomes in patients with endometriosis undergoing FET cycles, and determine the optimal number of GnRHa injections in GnRHa-HRT protocols. This was a retrospective cohort analysis of women with endometriosis who underwent FET cycles at a single university-based center. This study retrospectively analyzed 2048 FET cycles in our center from 2011 to 2020. According to the endometrial preparation protocols, patients were divided into 4 groups: gonadotropin releasing hormone agonist-hormone replacement therapy(GnRHa-HRT), hormone replacement therapy(HRT), ovulation induction(OI), and natural cycle(NC). In the GnRHa-HRT group, patients were further divided into 3 groups: one injection of GnRHa, two injections of GnRHa, and three or more injections of GnRHa. The primary outcome was the clinical pregnancy rate. Propensity score matching was used to adjust for potential non-similarities among the groups. Multivariate logistic regression analysis was performed to figure out the risk factors for pregnancy outcomes. There were no statistical differences in pregnancy outcomes among the four endometrial preparation protocols in FET cycles with endometriosis patients, the results retained after propensity score matching(PSM). And in endometriosis patients complicated with adenomyosis, the results remained similar. In patients with GnRHa-HRT protocol, there were no differences in clinical pregnancy rate and live birth rate with different numbers of GnRHa injections, the early miscarriage rate were 18% in the two injections of GnRHa group and 6.5% in the one injection of GnRHa group(P = 0.017). Multifactorial logistic regression analysis showed that two injections of GnRHa before FET was associated with increased early miscarriage rate compared with one injection of GnRHa[adjusted OR (95% CI): 3.116(1.079-8.998),p = 0.036]. The four kinds of endometrial preparation protocols for FET, GnRHa-HRT, HRT, OI and NC had similar pregnancy outcomes in patients with endometriosis. In endometriosis patients complicated with adenomyosis, the results remained similar. In patients with endometriosis undergoing GnRHa-HRT protocol for FET, more injections of GnRHa had no more advantages in pregnancy outcomes, on the contrary, it might increase the early miscarriage rate.


Smartphone Apps for Vaping Cessation: Quality Assessment and Content Analysis.
As the prevalence of electronic cigarette (e-cigarette) use, or vaping, continues to grow, particularly among young people, so does the need for research and interventions to address vaping. This study examines the quality of free vaping cessation apps, their contents and features, popularity among users, and adherence to evidence-based principles. A systematic search of existing apps for vaping cessation was conducted in December 2020. Eligible apps were free, in English, and included features specifically targeting vaping cessation. Each app included in the analysis was used daily for at least seven consecutive days, assessed using the Mobile App Rating Scale, and rated by at least two authors (AK, EL, or SS) based on adherence to evidence-based practices. Intraclass correlation coefficient (ICC) estimates were computed to assess interrater reliability (excellent agreement; ICC 0.92; 95% CI 0.78-0.98). A total of 8 apps were included in the quality assessment and content analysis: 3 were developed specifically for vaping cessation and 5 focused on smoking cessation while also claiming to address vaping cessation. The mean of app quality total scores was 3.66 out of 5. Existing vaping cessation apps employ similar approaches to smoking cessation apps. However, they are very low in number and have limited features developed specifically for vaping cessation. Given the lack of vaping cessation interventions at a time when they are urgently needed, smartphone apps are potentially valuable tools. Therefore, it is recommended that these apps apply evidence-based practices and undergo rigorous evaluations that can assess their quality, contents and features, and popularity among users. Through this process, we can improve our understanding of how apps can be effective in helping users quit vaping.


A study protocol of a three-group randomized feasibility trial of an online yoga intervention for mothers after stillbirth (The Mindful Health Study).
In the USA, stillbirth (in utero fetal death ≥20 weeks gestation) is a major public health issue. Women who experience stillbirth, compared to women with live birth, have a nearly sevenfold increased risk of a positive screen for post-traumatic stress disorder (PTSD) and a fourfold increased risk of depressive symptoms. Because the majority of women who have experienced the death of their baby become pregnant within 12-18 months and the lack of intervention studies conducted within this population, novel approaches targeting physical and mental health, specific to the needs of this population, are critical. Evidence suggests that yoga is efficacious, safe, acceptable, and cost-effective for improving mental health in a variety of populations, including pregnant and postpartum women. To date, there are no known studies examining online-streaming yoga as a strategy to help mothers cope with PTSD symptoms after stillbirth. The present study is a two-phase randomized controlled trial. Phase 1 will involve (1) an iterative design process to develop the online yoga prescription for phase 2 and (2) qualitative interviews to identify cultural barriers to recruitment in non-Caucasian women (i.e., predominately Hispanic and/or African American) who have experienced stillbirth (N = 5). Phase 2 is a three-group randomized feasibility trial with assessments at baseline, and at 12 and 20 weeks post-intervention. Ninety women who have experienced a stillbirth within 6 weeks to 24 months will be randomized into one of the following three arms for 12 weeks: (1) intervention low dose (LD) = 60 min/week online-streaming yoga (n = 30), (2) intervention moderate dose (MD) = 150 min/week online-streaming yoga (n = 30), or (3) stretch and tone control (STC) group = 60 min/week of stretching/toning exercises (n = 30). This study will explore the feasibility and acceptability of a 12-week, home-based, online-streamed yoga intervention, with varying doses among mothers after a stillbirth. If feasible, the findings from this study will inform a full-scale trial to determine the effectiveness of home-based online-streamed yoga to improve PTSD. Long-term, health care providers could use online yoga as a non-pharmaceutical, inexpensive resource for stillbirth aftercare. NCT02925481.


Ultrastructure of the cell wall and cytoplasmic membrane of gram-negative bacteria with different fixation techniques.
The ultrastructure of the cytoplasmic membrane and cell wall of two strains of Escherichia coli, Proteus morganii, P. vulgaris, Acinetobacter anitratum, Moraxella lacunata, Erwinia amylovora, Acinetobacter sp., and of a plant pathogen, unclassified gram-negative, fixed by the Ryter-Kellenberger procedure, was found to be significantly affected by the use or omission of the uranyl postfixation included in that procedure, and by the presence or absence of calcium in the OsO(4) fixative. The omission of the uranyl treatment results in a less clear profile of both the outer membrane of the cell wall and of the cytoplasmic membrane. The observation of these two membranes is further limited when both uranyl and calcium are omitted. The R-layer and the material covering the surface of the cell wall appear more distinct when the uranyl postfixation is not used. Evidence is given suggesting that the influence of uranyl and calcium ions on the appearance of the outer and cytoplasmic membranes would be primarily due to their action as fixatives, whereas the influence of uranyl on the appearance of the R-layer would be due to a direct action on the peptidoglycan component of this layer. When uranyl acetate is used as a section stain after the embedding in plastic, it improves the observation of the R-layer. In this case, a well contrasted R-layer is consistently observed in all strains studied, provided that the postfixation has been omitted. The frequent difficulty in clearly observing the R-layer in many published micrographs probably results from the common use of uranyl postfixation.