Seasonal dynamics of the microbiome-host response to pharmaceuticals and pesticides in Mytilus galloprovincialis farmed in the Northwestern Adriatic Sea.
Marine mussels, especially Mytilus galloprovincialis, are well-established sentinel species, being naturally resistant to the exposure to multiple xenobiotics of natural and anthropogenic origin. Even if the response to multiple xenobiotic exposure is well known at the host level, the role of the mussel-associated microbiome in the animal response to environmental pollution is poorly explored, despite its potential in xenobiotic detoxification and its important role in host development, protection, and adaptation. Here, we characterized the microbiome-host integrative response of M. galloprovincialis in a real-world setting, involving exposure to a complex pattern of emerging pollutants, as occurs in the Northwestern Adriatic Sea. A total of 387 mussel individuals from 3 commercial farms, spanning about 200 km along the Northwestern Adriatic coast, and in 3 different seasons, were collected. Multiresidue analysis (for quantitative xenobiotic determination), transcriptomics (for host physiological response), and metagenomics (for host-associated microbial taxonomical and functional features) analyses were performed on the digestive glands. According to our findings, M. galloprovincialis responds to the presence of the complex pattern of multiple emerging pollutants - including the antibiotics sulfamethoxazole, erythromycin, and tetracycline, the herbicides atrazine and metolachlor, and the insecticide N,N-diethyl-m-toluamide - integrating host defense mechanisms, e.g., through upregulation of transcripts involved in animal metabolic activity, and microbiome-mediated detoxification functions, including microbial functionalities involved in multidrug or tetracycline resistance. Overall, our data highlight the importance of the mussel-associated microbiome as a strategic player for the orchestration of resistance to the multixenobiotic exposure at the holobiont level, providing strategic functionalities for the detoxification of multiple xenobiotic substances, as occurring in real world exposure settings. Complementing the host with microbiome-dependent xenobiotic degradative and resistance genes, the M. galloprovincialis digestive gland associated microbiome can have an important role in the detoxification of emerging pollutants in a context of high anthropogenic pressure, supporting the relevance of mussel systems as potential animal-based bioremediation tool.


Hemodynamics at rest and during supine and sitting bicycle exercise in patients with coronary artery disease.
To access left ventricular function and compare pulmonary capillary wedge pressure and left ventricular end-diastolic pressure in the supine and sitting positions, 20 men with angina pectoris secondary to coronary artery disease underwent hemodynamic studies at rest and during exercise in the two positions. At rest the values for cardiac index, stroke index, systolic ejection rate index and left ventricular stroke work index were lower in the sitting position; heart rate, left ventricular end-diastolic pressure and pulmonary capillary wedge pressure were similar in the two positions. All patients experienced angina during both exercise periods. At angina during supine exercise, stroke index, systolic ejection rate index and left ventricular stroke work index did not increase significantly from the resting values. In contrast, during sitting exercise, significant increases in these variables were observed. Comparison of data during exercise revealed higher values for heart rate, mean systemic pressure, cardiac index, systolic ejection rate index, left ventricular stroke work index and rate-pressure product and lower values for mean pulmonary capillary wedge pressure (20 +/- 3 versus 27 +/- 3 [mean +/- standard error of the mean] mm Hg, P is less than 0.001), and left ventricular end-diastolic pressure (24+/- 3 versus 31 +/- 3 mm Hg, P is less than 0.02) in the sitting position; stroke index and S-T segment depression were similar during the two exercise periods. Four patients had insignificant increases in left ventricular filling pressure during both exercise periods. Of the 16 patients with abnormal left ventricular filling pressure during supine exercise, only 10 had a similar response during exercise in the sitting position. There was a good correlation between left ventricular end-diastolic pressure and mean pulmonary capillary wedge pressure at rest and during exercise in the two postures.


An inverse PCR technique to rapidly isolate the flanking DNA of dictyostelium insertion mutants.
Restriction enzyme mediated integration is a widely used and effective method for insertional mutagenesis in Dictyostelium discoideum. In this method, plasmid rescue is used to clone the genomic deoxyribonucleic acid (DNA) sequences that flank the insertion site. For this to be effective, it is necessary to first find a convenient restriction enzyme site within the genomic DNA. This is a time-consuming process that requires Southern blot analysis of the mutant DNA. In addition, plasmid rescue requires transformation into highly competent Escherichia coli. Problems can arise owing to unstable genomic sequences, damage to the plasmid DNA and exogenous plasmid contamination. We have established a simple and rapid polymerase chain reaction-based technique that works for all mutants and circumvents the need for Southern blot analysis and plasmid rescue.


Biomechanical Comparison of 3 Medial Patellofemoral Complex Reconstruction Techniques Shows Medial Overconstraint but No Significant Difference in Patella Lateralization and Contact Pressure.
The purpose of this study was to investigate biomechanical differences of medial patellofemoral ligament (MPFL) reconstruction, medial quadriceps tendon femoral ligament (MQTFL) reconstruction, and a combination of these techniques to restore lateral patellar constraint and contact pressures. Eight fresh frozen cadaver knees were mounted to a custom jig with physiological quadriceps tendon loading. Flexion angles and contact pressure (CP) were dynamically measured using Tekscan® pressure sensors and Polhemus® Liberty 6 degree of freedom (6DOF) positioning sensors in the following conditions: 1) intact 2) MPFL and MQTFL deficient, 3) MPFL reconstructed, 4) Combined MPFL + MQTFL reconstructed, and 5) MQTFL reconstructed. Lateral patellar translation was tested using horizontally directed 30 N force applied at 30° of knee flexion. The knees were flexed in dynamic fashion, and CP values were recorded for 10°, 20°, 30°, 50°, 70°, and 90° degrees of flexion. Group differences were assessed with ANOVA's followed by pairwise comparisons with Bonferroni correction. MPFL (P = .002) and combined MPFL/MQTFL (P = .034) reconstruction significantly reduced patellar lateralization from +19.28% (9.78%, 28.78%) in the deficient condition to -17.57% (-27.84%, -7.29%) and -15.56% (-33.61%, 2.30%), respectively. MPFL reconstruction was most restrictive and MQTFL reconstruction the least -7.29% (-22.01%, 7.45%). No significant differences were found between the three reconstruction techniques. Differences in CP between the three reconstruction techniques were not significant (<.02 MPa) at all flexion angles. The present study found no significant difference for patellar lateralization and patellofemoral CP between MPFL, combined MPFL/MQTFL, and MQTFL reconstruction. All 3 techniques resulted in stronger lateral patellar constraint compared to the native state, while the MQTFL reconstruction emulated the intact state the closest. Various surgical techniques for medial patellofemoral complex reconstruction can restore patellar stability with similar patellofemoral articular pressures.


[Wegener's granulomatosis with anti-neutrophil cytoplasmic antibodies against anti-cathepsin G antigen].
Wegener's granulomatosis belongs to the group of small vessel vasculitis associated with anti-neutrophil cytoplasmic antibodies characterized by granulomatous inflammation and necrotising vasculitis in various organs with particular involvement of the upper and lower respiratory tracts and kidneys. Wegener's granulomatosis is a rare disorder in childhood and early diagnosis of this disease is critical to the long-term prognosis of the disease. The presence of positive cytoplasmic antineutrophil cytoplasmic antibody staining or a high titre of proteinase 3 antibodies were added as new criteria of vasculitis in childhood. This article presents a case of Wegener's granulomatosis, with the presence of anti-neutrophil cytoplasm antibodies with cytoplasmic pattern with absence of anti-proteinase 3 antibodies and presence of high levels of anti-cathepsin G antibodies, rarely described in Wegener's granulomatosis.


Equilibrium and Kinetic Study of Lead and Copper Ion Adsorption on Chitosan-Grafted-Polyacrylic Acid Synthesized by Surface Initiated Atomic Transfer Polymerization.
In this work, we synthesized chitosan grafted-polyacrylic acid (CS-g-PA) through surface-initiated atom transfer radical polymerization (SI-ATRP). We also studied the adsorption process of copper and lead ions onto the CS-g-PA surface. Adsorption equilibrium studies indicated that pH 4.0 was the best pH for the adsorption process and the maximum adsorption capacity over CS-g-PA for Pb2+ ions was 98 mg·g-1 and for Cu2+ it was 164 mg·g-1, while for chitosan alone (CS), the Pb2+ adsorption capacity was only 14.8 mg·g-1 and for Cu2+ it was 140 mg·g-1. Furthermore, the adsorption studies indicated that Langmuir model describes all the experimental data and besides, pseudo-second-order model was suitable to describe kinetic results for the adsorption process, demonstrating a larger kinetic constant of the process was larger for Pb2+ than Cu2+. Compared to other adsorbents reported, CS-g-PA had comparable or even superior adsorbent capacity and besides, all these results suggest that the new CS-g-PA polymers had potential as an adsorbent for hazardous and toxic metal ions produced by different industries.


Prostacyclin and thromboxane synthesis by endometrial cancer and leiomyomas.
To study the role of prostacyclin (PGI2) and thromboxane A2 (TxA2) in uterine tumors, pieces of endometrial cancer (n = 12) and leiomyomas (n = 12) were incubated in vitro, and the productions of 6-keto-prostaglandin F1a (6-keto-PGF1a, a hydration product of PGI2) and thromboxane B2 (TxB2, a hydration product of TxA2), measured by radioimmunoassay, were compared to those of corresponding healthy tissues. The production of 6-keto-PGF1a by endometrial cancer (20.8; 15.1-85.0 ng/mg protein/min, median and interquartile range), by healthy endometrium (25.5; 10.0-55.0), by healthy myometrium (34.9; 25.0-59.9) and by leiomyoma (20.3; 10.2-45.1) was similar. The production of TxB2 was increased by endometrial cancer (55.5; 10.5-155.2, p less than 0.02) in comparison with endometrium (9.8; 4.3-35.1), myometrium (3.8; 2.1-8.0) and leiomyoma (1.9; 1.0-3.8). The 6-keto-PGF1a/TxB2 ratio in endometrial cancer (0.9; 0.3-1.5) was smaller (p less than 0.02) than that in healthy endometrium (3.3; 1.9-4.8). Thus, TxA2 may be a factor in endometrial cancer.


Disordered Eating and Body Dissatisfaction in Transgender and Gender-Expansive Adults: An Evaluation and Integration of the Gender Minority Stress and Resilience and Tripartite Influence Models.
Transgender and gender-expansive (TGE) people, including transgender and nonbinary individuals, experience elevated rates of body dissatisfaction and disordered eating compared to cisgender individuals, but little is known about why. To address this research gap, we compared the ability of TGE-specific factors as proposed in the gender minority stress and resilience (GMSR) model, general psychological factors contained in the tripartite influence (TI) model, and an integration of these frameworks to explain body dissatisfaction and disordered eating among TGE individuals. Regression analyses were conducted to test the models' abilities to explain experiences reported in a survey of 93 U.S. TGE adults, including 43 transgender women respondents (46.2%) and 31 transgender men respondents (33.3%). Participants were diverse with regard to age (M = 34.19, SD = 12.02) and ethnoracial background (e.g., 29.0% Hispanic/Latinx, 17.2% Black/African American, 6.5% American Indian/Alaska Native; 5.4% multiracial). Results demonstrated both models' abilities to explain body dissatisfaction and disordered eating independently, except for the relationship between body dissatisfaction and the thinness-oriented TI model. An integration of the models better explained body dissatisfaction and disordered eating compared to either model alone. Notably, some findings did not align with the two frameworks, suggesting existing models may not adequately describe pathways through which disordered eating emerges in TGE populations. Specifically, body dissatisfaction showed no significant relationship with disordered eating and was not well explained by the TI model, and higher gender identity pride was related to greater disordered eating symptoms. Implications for clinical care and future research are discussed.


Synthesis of long-chain alkyl glucosides via reverse hydrolysis reactions catalyzed by an engineered β-glucosidase.
Long-chain alkyl glucosides, such as octyl and decyl β-d-glucopyranosides (OG and DG, respectively), are regarded as a new generation of biodegradable, non-ionic surfactants. Previously, the mutants of Dalbergia cochinchinensis Pierre dalcochinase showed potential in the synthesis of oligosaccharides and alkyl glucosides. In this study, the N189F dalcochinase mutant gave the highest yields of OG and DG synthesis under reverse hydrolysis conditions. The optimized yield of OG (57.5 mol%) was obtained in the reactions containing 0.25 M glucose and 0.3 units of the N189 F mutant in buffer-saturated octanol at 30 °C. The identity of OG and DG products was confirmed by high resolution mass spectrometry (HRMS) and NMR. Consistent with its capability for synthesis, the reactivation kinetics and ITC analysis revealed that the aglycone binding pocket of the N189F mutant was more favorable for long-chain alkyl alcohols than the wild-type dalcochinase, while their glycone binding pockets showed similar affinity for the glucosyl moiety. STD NMR revealed higher interactions at the aglycone sites than the glycone sites. Our results demonstrated a promising potential of the N189F dalcochinase mutant in the future commercial production of long-chain alkyl glucosides via reverse hydrolysis reactions.


Cardiopulmonary resuscitation and acute cardiovascular life support--a protocol review of the updated guidelines.
For the first time in 5 years, new guidelines for cardiopulmonary resuscitation (CPR) of adults and children were introduced at the end of November 2005. The new CPR guidelines evolved from emerging evidence-based resuscitation studies and the evaluation process included the input of 281 international resuscitation experts who evaluated hypotheses, topics, and research over a 36-month period. The process included evidence evaluation, review of the literature, and focused analysis. This article reviews the four major changes to the guidelines. Changes are currently being made in the training of all new and recertifying ACLS health care providers.


The influence of Atorvastatin on ultrastructure of rat pancreatic exocrine cells in the course of experimental intoxication with ethanol.
The experiment was carried out on mature Wistar male rats. The animals from experimental group I received Atorvastatin in the dose 0.28 mg/24 h for 3 weeks. Experimental group II received 20% ethanol ad libitum apart from Atorvastatin. Ultrastructural examinations of pancreatic exocrine cells showed the stimulation of secretory processes in cells of animals receiving Atorvastatin and considerable damage of cell organelles in experimental group II, more intense than in the pancreas of control animals which drank ethanol.


Membrane-bound mRNAs are recruited from preinitiated ribonucleoprotein particles in injected Xenopus oocytes.
Messenger RNA injected Xenopus oocytes exhibit a differential capacity for translation. mRNAs translated in the free cytoplasm are translated efficiently whereas mRNAs translated on the rough endoplasmic reticulum (RER) membrane are translated inefficiently. If mRNA injected oocytes are injected additionally with proteins isolated from the RER, enhanced translation of RER-bound mRNAs is observed. When examined by sucrose gradient centrifugation and RNA dot blots, most of the injected RER-bound mRNA sediments less than or equal to the 80 S monosome. The RER proteins recruit these preinitiated mRNAs onto polysomes as evidenced by a shift in sedimentation to the polysome region of a sucrose gradient. When examined by immunoblotting, the RER proteins are shown to contain a protein which reacts specifically with an antibody directed against docking protein (SRP-receptor protein). However, this putative docking protein does not appear to be the protein which actually recruits the preinitiated mRNAs onto polysomes.


The impact of COVID-19 lockdown on the general health status of people with chronic health conditions in Belgium: a cross-sectional survey study.
Patients with chronic health conditions risk aggravation of their health status due to reduced access to health services during the COVID-19 related lockdown. To investigate the impact of Belgian COVID-19 measures on general health status (i.e. worse or stable/better) of patients, adult and pediatric, with chronic health conditions and how this change in health status relates to personal and health behavior-related factors. A cross-sectional study using an online survey was conducted during the first COVID-19 related lockdown in Belgium. Associations between change in health status since the lockdown and (change in) personal and health behavior-related factors (including physical activity, access to health-care services and social activities) were investigated. In adults (n = 561), almost all personal factors, including feelings of distress, depression, anxiety, somatization, and low self-efficacy, were significantly worse in patients with a worse health status during the lockdown (n = 293, 52%) compared to patients reporting a stable/better health status (p < .001-0.002). Also, these patients reported lower physical activity levels, more tele-consultations and less social activities (p < .001-0.006). In children (n = 55), all surveys were completed by a proxy (parent(s)/guardian) who reported a worse health status in 38% of the children. Level of distress of the child (p = .005) since the lockdown and somatization of the parent(s) (p = .0018) were significantly worse in children with a worse versus a stable/better health status. Fifty-two percent of the adults and 38% of children with chronic health conditions reported worsening of their general health status during the lockdown in March-May 2020 in Belgium. Negative personal factors and unhelpful health behavior seems to be associated with a worse health status.


No relevant influence on overall survival time in patients with metastatic breast cancer undergoing combination chemotherapy.
The results of clinical studies dealing with first and second line chemotherapy of metastatic breast cancer published between 1975 and early 1986 which involved 9350 women were reviewed. Our special aim was to evaluate combination chemotherapy and its influence on overall survival in late stage breast cancer patients. No significant improvement in overall survival times was found in this selected group of patients who were treated with intense palliative chemotherapy.


Evaluation of a comprehensive intervention with a behavioural modification strategy for childhood obesity prevention: a nonrandomized cluster controlled trial.
With regard to the global childhood obesity epidemic, it is imperative that effective lifestyle interventions are devised to combat childhood obesity. This paper describes the development and implementation of a comprehensive (a combination of diet and physical activity (PA)), social cognitive behaviour modification intervention using accelerometry and a dietary diary to tackle child overweight and obesity. The comprehensive intervention effect was evaluated in a comparison with diet only, PA only and a no-treatment control group. A pilot study was conducted with a non-randomized cluster design. Four hundred thirty-eight overweight and obese children aged 7-12 years from ten primary schools in Beijing were recruited to receive a one-year intervention. Participants were allocated into one of four groups: the comprehensive intervention group; the PA only group (Happy 10 program); the diet only group (nutrition education program); and a control group. The effects of intervention on adiposity, blood pressure, and biochemical indicators were assessed by examining 2-way interactions (time × intervention) in linear mixed models. Means and 95 % confidence intervals (CI) for the adjusted changes between post-intervention and baseline relative to changes in the control group were calculated and reported as effect sizes. The percentage of body fat in the comprehensive intervention group showed a significant relative decrease (adjusted change: -1.01 %, 95 % CI: (-1.81, -0.20) %) compared with the PA only, diet only or control groups (P < 0.001). Systolic blood pressure significantly decreased in the comprehensive intervention group (adjusted change: -4.37 mmHg, 95 % CI: (-8.42, -0.33) mmHg), as did diastolic blood pressure (adjusted change: -5.50 mmHg, 95 % CI (-8.81, -2.19) mmHg) (P < 0.05). Compared with the other two intervention groups and the control group, positive adjusted changes in fasting glucose in the comprehensive group were found, although not for the biochemical lipid metabolism indicators. Positive but non-significant adjusted changes in body mass index and waist circumference were observed. Compared with the diet or PA only intervention groups, the current comprehensive program had superior positive effects on body fat percentage and blood pressure but not on the biochemical lipid metabolism indicators in Chinese overweight and obese children. Future randomized controlled trials and long-term follow-up studies are required to elaborate the findings of the current intervention. ClinicalTrials.gov identifier: NCT02228434.


The associated burden of mental health conditions in alopecia areata: a population-based study in UK primary care.
Alopecia areata (AA) is a common cause of nonscarring hair loss that can have a profound psychological impact. To assess the co-occurrence of depression and anxiety in adults with AA compared with the general population, and to evaluate the mental health treatment burden and impact on time off work and unemployment. In total, 5435 people with newly diagnosed AA in UK primary care were identified from the Oxford Royal College of General Practitioners Research and Surveillance Centre network database, and matched to 21 740 controls. In cases and controls, we compared the prevalence and incidence of depressive episodes, recurrent depressive disorder and anxiety disorder, rates of time off work and unemployment, and, in those with pre-existing mental health conditions, rates of mental health-related prescribing and referral rates. This observational was registered with ClinicalTrials.gov (NCT04239521). Depression and anxiety were more prevalent in people diagnosed with AA than in controls (P < 0·001). People with AA were also more likely to subsequently develop new-onset depression and anxiety: adjusted hazard ratio (aHR) for recurrent depressive disorder 1·38 [95% confidence interval (CI) 1·13-1·69], depressive episodes aHR 1·30 (95% CI 1·04-1·62) and anxiety disorder aHR 1·33 (95% CI 1·09-1·63); to be issued time off work certificates (aHR 1·56, 95% CI 1·43-1·71); and to be recorded as unemployed (aHR 1·82, 95% CI 1·33-2·49). Higher rates of antidepressant prescribing were also seen in people with AA. People with AA have higher rates of depression and anxiety than those without AA. This impacts deleteriously on mental health treatment burden, time off work and unemployment. Evidence-based mental health treatment programmes are needed for people with AA.


Comprehensive analysis of chemokines family and related regulatory ceRNA network in lung adenocarcinoma.
Lung adenocarcinoma (LUAD) is one of the world's commonest malignancies with a high fatality rate. Chemokines not only regulate immune response but also participate in tumor development and metastasis and yet the mechanism of chemokines in LUAD remains unclear. In this study, transcriptional expression profiles, mutation data, and copy number variation data were downloaded from The Cancer Genome Atlas (TCGA). Risk gene protein expression was assessed by the Clinical Proteomic Tumor Analysis Consortium (CPTAC) and the Human Protein Atlas (HPA). Gene Expression Omnibus (GEO) data was used to validate the prognostic model. We summarized the genetic mutation variation landscape of chemokines. The risk prognosis model was developed based on differentially expressed chemokines, and patients in the high-risk score (RS) group had lower survival rates. Gene Set Enrichment Analysis (GSEA) revealed that high-RS patients were associated with metabolic transformation pathways, while low-RS patients were associated with immune-related pathways. Compared with the high-RS group, the low-RS group had higher immune/stromal/estimate scores calculated by the ESTIMATE package. The proportion of immune cells obtained using the CIBERSORT package was significantly different between the two groups. Most of the immune checkpoints were highly expressed in low-RS samples. Finally, we discovered that the lncRNA MIR17HG/AC009299.3/miR-21-5p/CCL20 regulatory network might be crucial in the pathogenesis of LUAD. In conclusion, we developed a risk signature and chemokine-related competing endogenous RNA (ceRNA) network.


Takayasu's disease presenting with atherothrombotic ischaemic stroke.
Takayasu's arteritis is a chronic inflammatory disease of the aorta and its main branches, and a well known cause of stroke. Pathogenesis of ischaemic stroke has been attributed to intracranial vasculitic involvement or emboli from either stenoocclusive extracranial vessels or cardiac disease such as aortic regurgitation. We present a patient with Takayasu's arteritis and recurrent cerebral infarctions associated with intracranial atherosclerosis. We postulate that the intracranial atherosclerotic process is an important mechanism in Takayasu's arteritis-related ischaemic stroke.


Thirty-Year Trends of Survival and Time-Varying Effects of Prognostic Factors in Patients With Metastatic Breast Cancer-A Single Institution Experience.
Metastatic breast cancer is generally considered an incurable disease. In our study we aimed to detect a time trend of survival over the past 30 years and account for time-varying effects of the prognostic factors. A total of 446 patients diagnosed with breast cancer at Saint Vincent de Paul Hospital, Lille, France between 1977 and 2013 who developed metastatic disease after a disease-free interval longer than 3 months and were followed-up for outcome. Data were analyzed using the Cox proportional hazards model and presented as hazard ratios (HRs). A monotonic time trend of survival was detected: a 2.6% lower risk of death for each increasing year over the past 30 years. Three prognostic factors had time-varying effects; the liver first metastasis (HR during the first 16 months of follow-up: 2.26; 95% confidence interval [CI], 1.65-3.11), the bone first metastasis (HR during the first 24 months of follow-up: 0.56; 95% CI, 0.43-0.74), and the disease-free interval (HR during the first 16 months of follow-up: 0.90; 95% CI, 0.85-0.95). The brain first metastasis, multiple first metastases, the lymph node ratio, and estrogen receptor status had a constant effect over time. In our study we detected a constant time trend of improvement in prognosis of metastatic breast cancer patients over the past 30 years and identified prognostic factors with time-varying effects.


Placental secretion of oestrogens and protein hormones.
In order to trace the passage of steroids and proteins from the placenta to the maternal peripheral blood, samples of retroplacental blood, uterine vein blood and peripheral blood were taken from women undergoing Caesarean section at term. The concentration of unconjugated oestriol, total oestriol, human placental lactogen and pregnancy specific beta1 glycoprotein was measured. The oestrogen concentration in the uterine vein was higher than the peripheral blood, the placental lactogen was slightly higher and the pregnancy specific beta1 glycoprotein was the same in the uterine vein and the peripheral veins. In keeping with this, the retroplacental blood had a high concentration of oestrogen, moderately raised placental lactogen and a low pregnancy specific glycoprotein concentration. It is concluded that there is a rapid secretion of oestriol from placenta to the mother; a slower secretion of placental lactogen and no clear evidence of a flow of pregnancy specific glycoprotein from the placenta.


Absence of fetal movements and normal infants.
Pregnant women feel movements from about the fifth month of pregnancy. In cases of high-risk pregnancy daily recording of fetal movements is useful in monitoring fetal well-being. Complete cessation of fetal movements points to a severely distressed fetus and impending death. We report here five women in whom failure to experience fetal movements was present from the beginning of pregnancy, or occurred after fetal movements had been felt for several weeks during the pregnancy. All the women delivered normal infants with high Apgar score.


Evidence of an association between the O blood group and allergic rhinitis.
The aim of this study was to verify if ABO phenotypes are associated with allergic rhinitis. 168 patients with allergic rhinitis and 168 control individuals from the same geographical region and paired by gender and age were enrolled in the study. ABO phenotypes were identified in red blood cells using the hemagglutination technique. The Fisher exact and chi-squared tests were employed to compare proportions. Statistical significance was set for an alpha error of 5% (p-value < 0.05). The overall differences in the frequencies of the ABO phenotypes of patients and controls were marginal (χ(2): 7.569; degrees of freedom (DF): 3; p-value = 0.055) however the O blood group was associated with allergic rhinitis (χ(2): 5.764; DF: 1; p-value = 0.016; OR: 1.735; CI 95%: 1.127-2.673). The differences in the frequencies of the O phenotype in patients and controls were statistically different for men (χ(2): 8.520; DF: 1; p-value = 0.003) but not for women (χ(2): 0.6375; DF: 1; p-value = 0.4246). The A phenotype was associated with protection (OR: 0.4385; CI 95%: 0.2043-0.9415; p-value = 0.049) and the O phenotype was associated with susceptibility (OR = 2.789; CI 95%: 1.385-5.616; p-value = 0.005) to allergic rhinitis only for men. The O blood group phenotype is associated with allergic rhinitis in male but not in female patients.


Thenar motor neuropathy electrophysiological study of 28 cases.
To report an electrophysiological study on thenar motor neuropathy of the median nerve. Twenty-eight consecutive patients (mean age, 48.8 years; 17 men) with dominant hand thenar muscle weakness without sensory symptoms were enrolled in this study. Electromyography of hand and forearm muscles and neurography of median, ulnar, radial, and palmar nerves, including distal motor latency recording from the second interosseous-lumbrical muscles, were performed. Complete denervation of the abductor pollicis brevis muscle was observed in one case and delayed median abductor pollicis brevis-distal motor latency was observed in the others. Other neurographic findings were normal. Thenar motor neuropathy may have different pathogeneses. It may be considered a variant of carpal tunnel syndrome involving the motor branch only or more likely due to chronic direct compression of the branch, because it preferentially affects males, dominant hand, and persons doing manual work. In both cases, anatomic origin and variations in the course of the branch may favor thenar motor neuropathy.


Non-greeting rituals in clinical placements.
This study aims to generate knowledge about how clinical placements contribute to the creation of sustainable nursing education in Denmark. The article discusses how nursing students feel socially excluded when registered nurses do not recognise them through deviating from culturally appropriate greeting rituals. A constructivist grounded theory study of (n = 15) individual semi-structured interviews with nursing students in their first and third years of study. Nursing students feel awkward, alienated and uncomfortable when confronted with a culture where registered nurses do not greet them. They are met by the attitude that Bauman refers to as "appropriate indifference," which means that the registered nurses act as if they neither see nor hear them because they are considered faceless private failures. Nurses' non-greeting rituals in clinical placements counteract the creation of sustainable nursing education. The aim of this article is to contribute with knowledge about socially inappropriate non-greeting rituals and recommend friendly and socially sustainable education in clinical placements.


High accuracy of recombinant fusion protein early secretory antigenic target protein 6-culture filtrate protein 10 skin test for the detection of tuberculosis infection: a phase III, multi-centered, double-blind, hospital-based, randomized controlled trial.
To verify the diagnostic utility of recombinant fusion protein ESAT6-CPF10 (EC), a novel skin test reagent to detect Mycobacterium tuberculosis infection. A multi-centered, double-blind, randomized controlled trial was conducted from December 17, 2015, to March 2, 2018. Participants involved in this study included those with active tuberculosis (TB), suspected pulmonary TB, or non-TB pulmonary disease. Each participant received three tests simultaneously, TB-specific enzyme-linked immunospot assay (T-SPOT.TB), tuberculin skin test (TST), and EC skin test (ECST), and adverse events were reported. Diagnostic accuracy was analyzed using data from 1085 protocol-compliant participants. The sensitivities of the ECST, TST, and T-SPOT.TB were 91.2% (95% CI, 89.0-93.2%), 91.4% (95% CI, 89.1-93.3%), and 92.1% (95% CI, 89.9-93.9%), respectively. The specificities of the ECST (69.7%, 95% CI, 64.5-74.5%) and T-SPOT.TB (76.1%, 95% CI, 71.2-80.5%) were significantly higher than the TST (54.4%, 95% CI, 48.9-59.7%). The agreements between ECST and TST (kappa = 0.632) and between ECST and T-SPOT.TB (kappa = 0.780) were substantial. No severe adverse event was reported. The diagnostic performance of the ECST was close to the T-SPOT.TB assay in the detection of TB infection and indicated good potential for clinical application in common scenarios.


In vitro inhibition of murine B-cell tumor growth by MDP, MDP(D-D) and Vaccin is mediated by macrophages.
The effect of two muramyl dipeptides, N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP) and N-acetylmuramyl-D-alanyl-D-isoglutamine (MDP(D-D)), on the in vitro growth of two murine ascitic plasmacytomas, MOPC 173 and TEPC 15, and on an ascitic lymphoma cell line, ABPL2, was studied. The ability of the muramyl dipeptides to inhibit tumor cell growth was compared with a sonicated antigenic preparation of Mycobacterium vaccae (Vaccin), known to have macrophage stimulation activity. The growth of all three ascitic cell lines was inhibited by both muramyl dipeptides and Vaccin. Macrophage-depletion of the ascitic cell populations led to an increase in cell growth of TEPC 15 and MOPC 173, and a decrease in ABPL2. A reduction or loss of the inhibitory activity of the muramyl dipeptides or Vaccin was also observed, and no inhibitory activity was found when the tumor cell lines were cultured in vitro to render them macrophage-free. The inhibitory activity of MDP or MDP(D-D) was restored when purified ascitic macrophages were added to the in vitro cultured cell lines. It was demonstrated that a minimum number of macrophages were necessary for the expression of inhibitory activity. Indomethacin, a PG-synthetase inhibitor, was found to act in a synergistic manner with MDP and MDP(D-D) in inhibiting TEPC 15, but antagonized the effect of these two agents on ABPL2. The lymphoma cell line ABPL2 appeared to be the most sensitive to inhibition by MDP(D-D), a nonpyrogenic, adjuvant-inactive stereoisomer of MDP.


Infected palmar pits: a rare presentation of Gorlin's syndrome.
This paper presents a rare hand presentation of Gorlin's Syndrome, also known as Naevoid Basal Cell Carcinoma Syndrome, which has not been reported previously. This condition is an autosomal dominant, multisystem condition whose diagnosis is important to ensure further surveillance and treatment of features of the condition not routinely dealt with by hand surgeons. However, hand surgeons should be aware of this condition and those parts of the syndrome which may present to them.


Use of Bisphosphonates, Calcium and Vitamin D for Bone Demineralization in Patients with Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome: A Systematic Review and Meta-Analysis of Clinical Trials.
The present study performed a systematic review and meta-analysis of clinical trials using bisphosphonates for bone demineralization in human immunodeficiency virus (HIV) patients. A comprehensive literature search was performed from January 2004 to January 2020 considering the bone mineral density (BMD) of the lumbar spine (LS) as the main outcome. Out of 214 titles that met criteria, 9 studies fulfilled the selection criteria. A total of 394 patients were identified, and they were allocated into 2 groups: the intervention group (200 patients), to whom a combination of alendronate or zoledronate with calcium and vitamin D was administered; and control group (194 patients), to whom only calcium and vitamin D was administered. Clinical profile and indicators of bone metabolism of the participants were evaluated regarding effect size, homogeneity, and consistency. No substantial heterogeneity between the groups was found for the baseline variables, and there was high consistency to the main outcome. The meta-analysis shows a significant difference in post-treatment BMD, favoring the intervention over the control treatment. The intervention improved LS density up to 0.227 g/cm², raising the average to the levels of general population. Adverse effects related to intervention were fever immediately after zoledronate administration and gastrointestinal complaints during alendronate usage. Other adverse effects were barely reported and poorly connected to intervention by studies' authors, despite all of them have been successfully resolved. This study provides evidence that BMD post-treatment is better in HIV patients who used bisphosphonates combined with calcium and vitamin D.


Evaluation of the age-dependent dosing recommendations for the administration of daily tinzaparin in children with thrombosis.
Essentials The once-daily dosing of tinzaparin provides an advantage over other low molecular weight heparins. The recommended age-dependent doses of tinzaparin in children have not previously been validated. Once-daily administration of tinzaparin is a safe and effective treatment of childhood thrombosis. Recommended doses are appropriate but monitoring may be required due to inter-individual variation. Background The recommended starting doses of tinzaparin for the treatment of thrombosis in children have not previously been validated. There are few data to support the efficacy and safety of once-daily tinzaparin dosing in children with thrombosis. Objectives To investigate the use of tinzaparin for the treatment of childhood thrombosis, and to evaluate the age-dependent dosing recommendations and define outcomes in terms of efficacy and safety. Methods This was a retrospective cohort study of children aged 0 to < 16 years treated for thrombosis at a large teaching hospital in the UK between 2008 and 2015. Medical records were reviewed to evaluate tinzaparin dosing, anti-activated factor X (FXa) levels, and patient outcomes. Results Seventy-nine children were identified as having received tinzaparin. Dosing information was available for 57. Younger children required higher doses to reach a therapeutic level. The therapeutic dose requirement varied within age groups, supporting the use of anti-FXa monitoring. Over a median follow-up of 35 months, there were 13 (16%) bleeding episodes (two major; seven clinically relevant but non-major; and four minor). There were two (3%) recurrent episodes of thrombosis. Children were treated for a median duration of 3 months, and the majority (86%) remained on tinzaparin for the duration of their anticoagulant therapy. Conclusion Once-daily tinzaparin is a safe and effective treatment for childhood thrombosis, with rates of recurrence and bleeding similar to those for other anticoagulants used in children. The recommended starting doses are appropriate, but anti-FXa monitoring may be required, owing to interindividual variability in the therapeutic dose requirement.


Diepoxybutane induces caspase and p53-mediated apoptosis in human lymphoblasts.
Diepoxybutane (DEB) is the most potent metabolite of the environmental chemical 1,3-butadiene (BD), which is prevalent in petrochemical industrial areas. BD is a known mutagen and human carcinogen, and possesses multiorgan systems toxicity that includes bone marrow depletion, spleen, and thymus atrophy. Toxic effects of BD are mediated through its epoxy metabolites. In working towards elucidating the cellular and molecular mechanisms of BD toxicity, we investigated the ability of DEB to induce apoptosis in human lymphoblasts. DEB induced a concentration and exposure time-dependent apoptosis, which accounted for the DEB-induced loss of cell viability observed in TK6 lymphoblasts. The DEB-induced apoptosis was inhibited by inhibitors of caspases 3 and 9. The role of p53 in mediating the DEB-induced apoptosis was also investigated. DEB induced elevated p53 levels in direct correlation to the extent of DEB-induced apoptosis, as the concentration of DEB increased up to 5 microM. The extent of DEB-induced apoptosis was dramatically higher in TK6 lymphoblasts as compared to the genetically paired p53-deficient NH32 lymphoblasts under the same experimental conditions. Our results confirm and extend observations on the occurrence of apoptosis in DEB exposed cells, and demonstrate for the first time the elevation of p53 levels in human lymphoblasts in response to DEB exposure. In addition, our results demonstrate for the first time that DEB-induced apoptosis is mediated by caspases 3 and 9, as well as the p53 protein. It is possible that DEB-induced apoptosis may explain BD-induced bone marrow depletion, spleen and thymus atrophy in BD-exposed animals.


Neuroendocrine differentiation in pure type mammary mucinous carcinoma is associated with favorable histologic and immunohistochemical parameters.
Mucinous carcinoma of the breast is a specific good prognostic type malignancy occurring in elderly patients. Neuroendocrine differentiation has long been described in mucinous carcinoma, but the significance of such finding is uncertain. We evaluated the neuroendocrine differentiation profiles of 38 cases of pure mucinous carcinoma and compared the clinicopathological differences between those with and those without neuroendocrine differentiation. The parameters assessed included patients' age, tumor size, nuclear grade, axillary lymph node status at time of diagnosis, percentage area of intratumoral mucin, and the expression of estrogen and progesterone receptors, cerbB2 oncoprotein, and three neuroendocrine markers, namely neurone-specific enolase, chromogranin, and synaptophysin by immunohistochemistry. Patients' outcome and follow-up period were also documented. Of the 38 cases of pure mucinous carcinoma, 28, 11 and six cases showed positive staining for 1, 2 and 3 of the neuroendocrine markers. For all the groups with variable neuroendocrine differentiation and compared to those without such differentiation, they all showed older patients' age, higher proportion of tumors with lower nuclear grade, lower incidence of axillary lymph node metastasis, a higher progesterone receptor, and lower cerbB2 oncoprotein expression. No difference was detected between tumor size, intratumoral mucinous area, and estrogen receptor status. In all, 37 patients did not have distant metastases or local recurrences at the end of follow-up period, while one patient with coexisting high-grade ductal carcinoma in situ at time of diagnosis died of breast carcinoma. Our findings suggest that the identification of neuroendocrine differentiation in pure mucinous carcinoma is associated with more favorable histologic and immunohistochemical parameters.


Electric field effects in bacteriorhodopsin.
Exposure of aqueous suspensions of fragments of the purple membrane of Halobacterium halobium to electric field pulses leads to transient linear dichroism phenomena. The effects are interpreted in terms of field-induced alignments of the bacteriorhodopsin chromophore. Two observed relaxation times (tau) are attributed to rotation of the whole membrane fragments (tau s approximately 100 ms), and to a much faster reorientation of the chromophore within membrane (tau f approximately 260 microns).


Involvement of interleukin-1beta-converting enzyme in apoptosis of irradiated retinoblastomas.
To investigate whether interleukin-1beta-converting enzyme (ICE), a mammalian homologue of the Caenorhabditis elegans cell death gene ced-3, is involved in gamma-irradiation-induced apoptosis (programmed cell death) of human retinoblastoma cells. The induction of apoptotic cell death in human retinoblastoma cell lines WERI-Rb-1 and Y79 by gamma-irradiation was determined with a modified 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide colorimetric assay and the DNA-binding fluorochrome bis (benzimide) trihydro-chloride (Hoechst 33258) staining. The change of ICE protein level in tumor cells during apoptosis was determined by immunoblotting assay. Whether the specific tetrapeptide ICE inhibitor Ac-YVAD-CMK affected gamma-irradiation-induced apoptosis in tumor cells was also examined. The effect of ICE overexpression on tumor cells was evaluated by a transient transfection assay using ICE expression vector. Gamma-irradiation inhibited the cell viability of WERI-Rb-1 and Y79 cells in a dose-dependent manner and induced apoptosis. The protein level of ICE was remarkably enhanced after the treatment. The apoptotic cell death induced by gamma-irradiation was suppressed by the tetrapeptide ICE inhibitor Ac-YVAD-CMK. Moreover, overexpression of ICE induced apoptosis in tumor cells. These findings suggest that ICE may play an important role in gamma-irradiation-induced apoptosis in retinoblastoma cells. Transfer of the ICE gene induces apoptosis in these cells without gamma-irradiation.


Erratum to: Ultrapure dialysis water obtained with additional ultrafilter may reduce inflammation in patients on hemodialysis.
In original publication, the Table 4 was incorrect. The correct Table has been given below.


In vivo spectrometric calcium flux recordings of intrinsic Caudate-Putamen cells and transplanted IMR-32 neuroblastoma cells using miniature fiber optrodes in anesthetized and awake rats and monkeys.
A method is described to enable the recording of transient intracellular calcium changes in deep brain structures in anesthetized and awake animals using a fluorescent indicator combined with in vivo optical detection methods. Optrodes were fabricated using a bifurcated fiber-optic cable with an attached infusion guide cannula. After intracranial implantation of an optrode, animals were prepared in the following manner, (1) rats (intra-striatal) and monkeys (intra-putamen) were infused with the fluorescent calcium indicator, Oregon Green, to load intrinsic cells; or (2) rats were intra-striatally transplanted with a slurry of dye-loaded IMR-32 neuroblastoma cells via pipette ejection. Excitation light from an argon-ion laser was launched through the optrode and passed into the tissue. The resulting calcium-induced fluorescence signals were captured by the optrode, then detected and processed by externalized photomultiplier- and CCD-based spectrometer electronics. In approximately 25% of all intrinsic cell recordings, the baseline fluorescence intensity was relatively stable over time whereas in the remainder, large amplitude oscillations were observed with a frequency in the range of 0.5-2 Hz. These Ca(2+) transients were inhibited by local infusion of 10 microM omega-conotoxin MVIIC and 1 microM TTX. Extracellular electrophysiological recordings that were made adjacent to the optrode tip revealed that the Ca(2+) oscillations were in phase with the burst firing of striatal neurons. This suggested that the optical signals had a neuronal origin, most likely from medium spiny neurons. Baseline fluorescence intensity increased during infusion of high [K(+)](o), the calcium ionophore, A-23187, or during temporary bilateral carotid artery occlusion. Monkey (Saimiri sciureus) putamen recordings also affirmed the presence of similar calcium-related transients in a non-human primate. In the transplant preparations, the IMR-32 cells displayed a stable, non-oscillating baseline fluorescence. They were similarly responsive to high [K(+)](o) challenge and appeared viable for at least several hours. Similar optical recording approaches might be applied to monitor other fluorescent, chemiluminescent or bioluminescent events from almost any brain structure. Moreover, transplanted transfected cells expressing a single specific receptor or ion-channel protein may effectively serve as biosensing elements for the measurement of extracellular neurochemical signaling.


Chylothorax as a complication of harvesting the left internal thoracic artery in coronary artery bypass surgery.
A patient is described with chylothorax after harvesting the left internal thoracic artery for coronary artery bypass surgery. Conservative treatment with a medium chain triglycerides diet and tube thoracostomy was not only unsuccessful, but also resulted in the complications of malnutrition and empyema. We recommend early surgical treatment of chylothorax after coronary bypass surgery with the use of an internal thoracic artery.


Stability of Fentanyl Citrate, Hydromorphone Hydrochloride, Ketamine Hydrochloride, Midazolam, Morphine Sulfate, and Pentobarbital Sodium in Polypropylene Syringes.
Determine the stability of fentanyl 10 mcg/mL in 0.9% sodium chloride, fentanyl 10 mcg/mL in 5% dextrose, fentanyl 50 mcg/mL, hydromorphone 100 mcg/mL in 0.9% sodium chloride, ketamine 10 mg/mL, midazolam 0.4 mg/mL in 5% dextrose, midazolam 5 mg/mL, morphine 1 mg/mL in 0.9% sodium chloride, morphine 1 mg/mL in 5% dextrose, and pentobarbital 50 mg/mL when stored as single drug entities at room temperature in polypropylene syringes. Four 5 mL samples of each drug and concentration were prepared in 10 mL polypropylene syringes. The samples were stored at ambient room temperature in a locked cabinet. Triplicate determinations of drug concentration for each sample were performed initially, on day 50 or 51, and on day 100 using high-performance liquid chromatography with diode-array detection. With the exception of the hydromorphone 100 mcg/mL dilution, all compounds were found to contain greater than 95% of their initial concentration remaining at 100 days. Each sample remained clear and colorless when visually inspected.


Acute tumor lysis syndrome after proximal splenic artery embolization.
Preoperative splenic artery embolization for massive splenomegaly has been shown to reduce intraoperative hemorrhage during splenectomy. We describe a case of tumor lysis syndrome after proximal splenic artery embolization in a patient with advanced mantle cell lymphoma and splenic involvement. The patient presented initially with hyperkalemia two days after embolization that worsened during splenectomy. He was stabilized, but developed laboratory tumor lysis syndrome with renal failure and expired. High clinical suspicion of tumor lysis syndrome in this setting is advised. Treatment must be started early to avoid serious renal injury and death. Lastly, same day splenectomy and embolization should be considered to decrease the likelihood of developing tumor lysis syndrome.


Paraplegia Following Type B Acute Aortic Dissection Can Spare the Spinal Cord.
Ischemic lumbosacral plexopathy secondary to an acute aortic dissection is a rare condition that is usually unilateral and frequently accompanied by a simultaneous spinal cord infarction. The functional prognosis relies on the severity of the nervous system involvement being usually worse when the spinal cord is involved. We present a case of a 46-year-old man who suffered an acute type B aortic dissection presenting as acute paraplegia due to bilateral ischemic lumbosacral plexopathy treated with thoracic endovascular aortic repair. An up-to-date review of the literature on ischemic lumbosacral plexus injury is provided.


Stimulus properties of fluvoxamine in a conditioned taste aversion procedure.
A conditioned taste aversion (CTA) procedure in mice was used to investigate the stimulus effects of the serotonin reuptake inhibitors (SSRIs) fluvoxamine and fluoxetine. Fluvoxamine elicited a reliable CTA (ED50 = 24 mg/kg, SC) and a number of drugs were tested as pre-exposure drugs. Pre-exposure to the serotonin (5-HT)1A receptor agonists flesinoxan and +/- -8-hydroxy-dipropylaminotetralin (8-OH-DPAT) prevented the CTA induced by fluvoxamine (50 mg/kg, SC). Pre-exposure with the 5-HT2C receptor agonist MK 212 [6-chloro-2(1-piperazinyl)pyrazine] partially prevented the fluvoxamine-induced CTA, pre-exposure with the 5-HT2A/2C receptor agonist DOI [1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane HCl] did not prevent the CTA induced by fluvoxamine. Flesinoxan pre-exposure also prevented the taste aversion induced by fluoxetine (10 mg/kg, SC) completely. This contrasts previous results obtained with fluoxetine, where was found that its stimulus is primarily mediated by 5-HT2C, and to a lesser degree by 5-HT1A receptors. Therefore, we compared the two SSRIs directly. Pre-exposure to fluvoxamine prevented the fluoxetine-induced CTA, whereas pre-exposure to fluoxetine only partially prevented the fluvoxamine-induced CTA. We conclude that 5-HT1A receptors are involved in the stimulus properties of both fluvoxamine and fluoxetine, that 5-HT2C receptors are involved in fluvoxamine and especially fluoxetine, and, based primarily on the cross-comparison tests, that the two SSRIs have somewhat different stimulus properties.


The Influence of New York's SAFE Act on Individuals Seeking Mental Health Treatment.
Responding to mass shootings, some states have passed gun removal laws (e.g., NYS' SAFE Act), requiring that mental health professionals report patients who might potentially harm themselves/others. The purpose of this study was to assess whether knowledge of the SAFE Act impacts patients' mental health treatment-seeking and symptom-reporting behaviors. Patients at two mental health centers were surveyed during 2014-2018. Participants were asked if they would be concerned about being reported to county government, likelihood of seeking mental health treatment, and willingness to report mental health symptoms/behaviors given the SAFE Act's provisions. 228 patients (71.5% response rate) completed questionnaires. About 18% were concerned about being reported to county government, 9% would be less likely to seek mental health treatment, and about 23% would be less willing to report mental health symptoms/behaviors. Although these behaviors were not affected for most participants, there was a small minority who were concerned and less willing to report mental health symptoms/behaviors.


N-methyl-D-aspartate- and non-N-methyl-D-aspartate-evoked adenosine release from rat cortical slices: distinct purinergic sources and mechanisms of release.
Excitatory amino acids, acting at both N-methyl-D-aspartate (NMDA) and non-NMDA receptors, release the inhibitory neuromodulator adenosine from superfused rat cortical slices. This study was initiated to investigate the possible purinergic sources and mechanisms of release for the adenosine release evoked by NMDA and non-NMDA receptor activation. Inhibition of the bidirectional nucleoside transporter with dipyridamole greatly enhanced adenosine release evoked by glutamate. NMDA, kainate, and (RS)-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA). Inhibition of ecto-5'-nucleotidase with alpha, beta-methylene ADP and GMP had no effect on either kainate- or AMPA-evoked adenosine release, but it decreased glutamate- and NMDA-evoked adenosine release by 23 and 68%, respectively. A similar inhibition of NMDA-evoked adenosine release was observed with alpha, beta-methylene ADP alone, indicating that the inhibitory effect was not due to the reported competitive inhibition of NMDA receptors by GMP. Finally, NMDA-evoked adenosine release, but not kainate- or AMPA-evoked release, was Ca2+ dependent. These results indicate that activation of non-NMDA receptors releases adenosine per se in a Ca(2+)-independent manner. In contrast, NMDA receptor activation releases primarily a nucleotide that is subsequently converted extracellularly to adenosine; in this case, release is Ca2+ dependent. Although neither NMDA- nor non-NMDA-evoked adenosine release occurs via the nucleoside transporter, this transporter does appear to be a major route for removal of adenosine from the extracellular space.


Dietary exposure to continuous small doses of α-cypermethrin in the presence or absence of dietary curcumin does not induce oxidative stress in male Wistar rats.
α-Cypermethrin is a widely used insecticide and, at high doses, induces oxidative stress in mammals. Curcumin is an antioxidant phytochemical commonly used for food coloring and flavoring. We aimed to investigate the effects of continuous dietary exposure to low doses of α-cypermethrin, as is the case in exposed humans, on oxidative stress and its potential prevention by dietary curcumin. Four groups of ten male Wistar rats were ad libitum-fed a control diet or identical diets fortified with α-cypermethrin (350 mg/kg diet), curcumin (1000 mg/kg diet), or α-cypermethrin and curcumin (350 and 1000 mg/kg diet, respectively) for 7 weeks. α-Cypermethrin accumulated in adipose tissues and was detectable in kidney, liver, and brains. Dietary α-cypermethrin did not alter concentrations of malondialdehyde, ascorbic and uric acid, retinol, liver damage markers, or the activities of CAT and SOD, but reduced vitamin E in blood. α-Cypermethrin did not affect malondialdehyde or reduced glutathione concentrations in any of the tissues, but significantly increased glutathione disulfide in kidney and subcutaneous adipose tissue. In conclusion, dietary exposure to small doses of α-cypermethrin did not induce oxidative stress in rats and may be less toxic than exposure to comparable quantities administered as single high doses by gastric intubation.


Lipoprotein lipase and endothelial lipase expression in mouse brain: regional distribution and selective induction following kainic acid-induced lesion and focal cerebral ischemia.
Lipoprotein and endothelial lipases are members of the triglyceride lipase gene family. These genes are expressed in the brain, where the encoded proteins are fulfilling functions that have yet to be elucidated. In this study, we examined the distribution of their respective mRNAs in the C57BL/6 mouse brain by in situ hybridization. In control mice, we observed widespread expression of lipoprotein lipase (LPL) mRNA mainly in pyramidal cells of the hippocampus (CA1, CA2 and CA3 areas), in the striatum and in several cortical areas. Endothelial lipase (EL) mRNA expression was restricted to CA3 pyramidal cells of the hippocampus, to ependymal cells in the ventral part of the third ventricle and to some cortical cell layers. To gain insight into the role played by lipases in the brain, neurodegeneration was induced by intraperitoneal injection of kainic acid (KA) or by occlusion of the middle cerebral artery (MCA). Upon injection of KA, a rapid increase in EL mRNA expression was observed in the piriform cortex, hippocampus, thalamus and neocortex. However, the levels of LPL mRNA were unaffected by KA injection. Remarkably, after focal cerebral ischemia, the expression of EL was unaffected whereas a dramatic increase in LPL expression was observed in neocortical areas of the lesioned side of the brain. These results show that LPL and EL transcripts are selectively upregulated in function of the type of brain injury. LPL and EL could thus fulfill a function in the pathophysiological response of the brain to injury.


Umbilical artery flow velocity waveforms in monoamniotic twins with cord entanglement.
Monoamniotic twins with ultrasonographic evidence of umbilical cord entanglement present a management challenge in the antepartum period. We report two pregnancies with monoamniotic twins and ultrasonographic evidence of cord entanglement in which longitudinal Doppler flow velocity waveforms of the umbilical and middle cerebral arteries were obtained. A notch in the umbilical artery velocity waveform was noted in one twin member at 33 weeks' gestation and in both twin members of the other pregnancy at 31 weeks' gestation. Worsening of the umbilical artery velocity waveform notch with advancing gestation was noted in one fetus. The presence of a notch in the umbilical artery velocity waveform may reflect hemodynamic alterations in the fetal-placental circulation secondary to narrowing of the umbilical vessels involved in cord entanglement. This Doppler finding may be useful in the antepartum management of these pregnancies.


Dynamics of urine proteomics biomarker and disease progression in patients with IgA nephropathy.
Immunoglobulin A nephropathy (IgAN) frequently leads to kidney failure. The urinary proteomics-based classifier IgAN237 may predict disease progression at the time of kidney biopsy. We studied whether IgAN237 also predicts progression later in the course of IgAN. Urine from patients with biopsy-proven IgAN was analyzed using capillary electrophoresis-mass spectrometry at baseline (IgAN237-1, n = 103) and at follow-up (IgAN237-2, n = 89). Patients were categorized as "non-progressors" (IgAN237 ≤0.38) and "progressors" (IgAN237 >0.38). Estimated glomerular filtration rate (eGFR) and urinary albumin-creatinine ratio slopes were calculated. Median age at biopsy was 44 years, interval between biopsy and IgAN237-1 was 65 months and interval between IgAN237-1 and IgAN237-2 was 258 days (interquartile range 71-531). IgAN237-1 and IgAN237-2 values did not differ significantly and were correlated (rho = 0.44, P < .001). Twenty-eight percent and 26% of patients were progressors based on IgAN237-1 and IgAN237-2, respectively. IgAN237 inversely correlated with chronic eGFR slopes (rho = -0.278, P = .02 for score-1; rho = -0.409, P = .002 for score-2) and with ±180 days eGFR slopes (rho = -0.31, P = .009 and rho = -0.439, P = .001, respectively). The ±180 days eGFR slopes were worse for progressors than for non-progressors (median -5.98 versus -1.22 mL/min/1.73 m2 per year for IgAN237-1, P < .001; -3.02 vs 1.08 mL/min/1.73 m2 per year for IgAN237-2, P = .0047). In multiple regression analysis baseline progressor/non-progressor according to IgAN237 was an independent predictor of eGFR180days-slope (P = .001). The urinary IgAN237 classifier represents a risk stratification tool in IgAN also later in the course of the dynamic disease. It may guide patient management in an individualized manner.


MMP-2 serum concentrations predict mortality in hemodialysis patients: a 5-year cohort study.
We evaluated the ability of matrix metalloproteinase (MMP)-2, MMP-9, myeloperoxidase, osteopontin and stromal cell-derived factor 1 to predict mortality in hemodialysis (HD) patients. One hundred forty HD patients were enrolled and followed from December 2007 until December 2012. At the end of this 5-year period, data were compared between the patients who were alive and those who had died. The patients who alive were younger (56 vs. 63y), with lower frequency of diabetes mellitus (34.34% vs. 58.53%), higher concentrations of albumin (4.13 vs. 3.91mg/dl) and lower concentrations of MMP-2 (430.76 vs. 521.59ng/ml). Multivariate analysis showed that age (HR=1.03, p=0.02), diabetes mellitus (HR=2.395, p=0.012), albumin (HR=0.475, p=0.047) and MMP-2 (HR=1.003, p=0.005) were independent factors predicting mortality in HD patients. Receiver operating characteristic curve analysis showed that albumin (AUC=0.628, p=0.027) and MMP-2 (AUC=0.643, p=0.004) had a similar ability (p=0.76) to predict survival of HD patients. Compared with albumin, serum MMP-2 is a non-inferior prognostic marker for predicting the survival of HD patients.


Platelet rich plasma transforms exogenous prostaglandin endoperoxide H2 into thromboxane A2.
Platelet rich plasma transforms exogenous prostaglandin endoperoxide H2 into thromboxane A2 immediately prior to the initiation of irreversible aggregation. Selective thromboxane synthetase inhibitors block thromboxane A2 formation and aggregation. Thromboxane A2 formation appears to be essential during arachidonate mediated aggregation. The results presented reconcile the previously accepted paradoxical behavior of thromboxane synthetase in platelet rich plasma toward the prostaglandin endoperoxide H2 substrate.


Impact of polystyrene microplastics on major marine primary (phytoplankton) and secondary producers (copepod).
The effect of microplastic adsorption on marine microalgae Tetraselmis suecica, Amphora subtropica, and copepod Pseudodiaptomus annandalei was investigated in the present study. Fluorescence microscopic images were used to evaluate MP interactions with algae and copepods. T. suecica growth rate decreased with effects of 0.1 µm polystyrene exposure to 75 µl/100 ml (0.899 to 0.601 abs), 50 µl/100 ml (0.996 to 0.632 abs) and 25 µl/100 ml (0.996 to 0.632 abs), respectively. On the other hand, at 10th day of experiment, the control T. suecica showed the highest growth rate (0.965 abs), chlorophyll concentration (Chl-'a' = 21.36 µg/L; Chl-'b' = 13.65 µg/L), and cell density (3.3 × 106 cells/ml). A marine diatom A. subtropica absorbed 2.0 μm microplastics, and the maximal inhibition rate increased at higher MP concentration until 10th day. The highest MPs (75 μl/100 ml) treatment resulted in decreased growth rate of A. subtropica from 0.163 to 0.096 abs. A. subtropica (without MPs) had the highest lipid concentration of 27.15%, whereas T. suecica had the lowest lipid concentration of 11.2% (without MP). The maximum survival (80%) of P. annandalei was found in control on 15th day whereas on 12th day, the microplastics ingested copepod had the lowest survival rate (0%). On 15th day, the maximum Nauplii Production Rate (NPR) (19.33) female-1 was observed in control, whereas the minimum (17.33) female-1 NPR was observed in copepod ingested with MPs. The maximum lipid production (17.33% without MPs) was reported in control, whereas MPs fed copepods had the lowest lipid production (16%). Long-term exposure to polystyrene microplastics significantly reduced algae growth and chlorophyll concentration and also NPR and lipid concentration rate of copepod. We inferred that microplastic exposure of algae and copepods might results in persistent decreases in ingested carbon biomass over time.


Antioxidant action and photosensitizing effects of three different chlorpromazines.
Chlorpromazine inhibits by about 60% the lipid peroxidation stimulated by Fe2+/ascorbate in liposomes and the lipid peroxidation stimulated by cumene hydroperoxide in microsomes. Under the same conditions, two new synthetic derivatives of chlorpromazine, i.e., a N-benzoyloxymethylchlorpromazine and a N-pivaloyloxymethylchlorpromazine, induce no more than a 20% inhibition. On the other hand, when the different chlorpromazines are entrapped in liposomes and subsequently irradiated with near-UV light, they act as photosensitizing agents giving rise to lipid peroxidation. The latter is quite extensive in the presence of chlorpromazine or N-pivaloyloxymethylchlorpromazine, whereas it is drastically lower in the presence of N-benzoyloxymethylchlopromazine. The N-benzoyloxymethylchlorpromazine molecule, despite its low photodynamic effect, retains its neuroleptic properties. The possible mechanisms of the antioxidant and prooxidant actions of these compounds are discussed.


Association of serum bilirubin and uric acid levels changes during neuroinflammation in patients with initial and relapsed demyelination attacks.
In order to examine the endogenous antioxidants values in the earliest phase of demyelination, we have determined bilirubin and uric acid (UA) serum values in the patients with clinically isolated syndrome (CIS) and relapsing remitting multiple sclerosis (RRMS), regarding their clinical disability, measured by Extended Disability Status Scale (EDSS), Magnetic Resonance Imaging (MRI), disease duration, gender and other parameters. The bilirubin and UA levels were lower in CIS and RRMS patients than in control group, whether male or female (p < 0.05). The bilirubin and UA levels were decreased in RRMS compared to CIS patients (p < 0.05). Regarding EDSS, MRI and disease duration, obtained values of bilirubin and UA were higher in both study groups in patients with lower EDSS, lower MRI lesion number and shorter disease duration (p < 0.05). The greatest significance in decreased bilirubin and UA levels was observed in female compared to male patients, in both study groups (p < 0.05). The results suggest negative linear correlation between bilirubin and UA levels and disease duration, EDSS and MRI in CIS (p < 0.01), with the same correlation between bilirubin and UA levels and disease duration in RRMS patients (p < 0.01). There was also significant correlation between bilirubin level and MRI findings and UA levels and EDSS in RRMS patients (p < 0.01). The obtained results point to the importance of endogenous antioxidants in the outbreak and course of neuroinflammation. This could be favorable for the new pathogenetically conditioned neuroinflammatory therapy concepts which do not initially rely only on immunomodulatory, but also on the antioxidative effects.


Chk1/2 inhibitor AZD7762 blocks the growth of preantral follicles by inducing apoptosis, suppressing proliferation, and interfering with the cell cycle in granulosa cells.
Checkpoint kinases 1/2 (Chk1/2) have an important role in somatic cell development and oocyte meiotic maturation. However, the role of Chk1/2 in folliculo-genesis has not been fully elucidated. The aim of this study was to assess the effects of Chk1/2 inhibition on ovarian folliculogenesis and granulosa cell development in mice. Preantral follicles (100-120 μm) and granulosa cells from pre-ovulatory follicles (pre-GCs) of mice were isolated and cultured with or without Chk1/2 inhibitor AZD7762. Preantral follicles were cultured for 96h. Then, follicle morphology and fol-licular growth were assessed every 48h. Granulosa cells were cultured for 48h with or without AZD7762, after which cell apoptosis, cell proliferation, and cell cycle analysis were assessed; meanwhile, the mRNA expression of PCNA and Bax were measured by real-time RT-PCR, and PCNA and Bax protein were measured by Western blot. Compared with control follicles, AZD7762 inhibited growth of preantral folli-cles (P<0.05). Furthermore, inhibition of Chk1/2 significantly induced apoptosis (P<0.05) and inhibited the proliferation of granulosa cells (P<0.01), arrested cell cycle at S and G2/M phases, and decreased G1 phase fraction (P<0.001). Also, the expres-sion of PCNA mRNA and protein were reduced (P<0.01), while Bax mRNA and pro-tein were increased (P<0.05) post AZD7762 treatment in granulosa cells. This study revealed that Chk1 and Chk2 have a crucial role during preantral follicular development by regulating the proliferation and apoptosis of granu-losa cells.


Genetic association between IL23R rs11209026 and rs10889677 polymorphisms and risk of Crohn's disease and ulcerative colitis: evidence from 41 studies.
IL23R plays an important role in the pathogenesis of inflammatory bowel disease (IBD). The IL23R rs11209026 and rs10889677 polymorphisms have been shown to be associated with the development of Crohn's disease (CD) and ulcerative colitis (UC). But the results were inconsistent and inconclusive. So, we aim to investigate the genetic association between rs11209026 and rs10889677 polymorphisms and UC and CD risk by a meta-analysis. Literature search was conducted through PubMed, CNKI, and web of science databases. Pooled OR and 95% CI was used to assess the association between the allelic, dominant and recessive models of IL23R rs11209026 and rs10889677 polymorphisms and UC and CD risk. 41 publications with 13,803 patients with CD and 17,446 controls, as well as 5876 patients with UC and 10,053 controls were included in the present study. All the genetic models of rs11209026 polymorphism significantly decrease CD and UC risk (except for the recessive model in UC) (p < 0.05). A subgroup analysis based on ethnicity showed that the allelic (p < 0.00001, OR = 0.65) and dominant models (p < 0.00001, OR = 0.61) of rs11209026 polymorphism were significantly associated with UC risk in Caucasians, but not in Asians (p > 0.05). In addition, the allelic (CD: p < 0.00001, OR = 1.34; UC: p < 0.00001, OR = 1.22) and dominant models (CD: p = 0.002, OR = 1.39; UC: p = 0.01, OR = 1.29), but not the recessive model of rs10889677 polymorphism significantly increase the risk of CD and UC (p > 0.05). A subgroup analysis showed that the genetic models of rs10889677 polymorphism were associated with CD risk in Caucasians (p < 0.05), but not in Asians (p > 0.05). The dominant model of rs10889677 polymorphism was associated with UC risk in Asians (p = 0.04, OR = 1.54), but not in Caucasians (p > 0.05). Our meta-analysis demonstrated that the rs11209026 polymorphism might be a protective factor against developing IBD, while the rs10889677 polymorphism might be a risk factor for IBD.


Efficacy of tenofovir-based rescue therapy in patients with lamivudine-resistant hepatitis B virus: A systematic review and meta-analysis.
Currently, there are no conclusive results on the efficacy of Tenofovir disoproxil fumarate (TDF) monotherapy in chronic hepatitis B (CHB) patients with lamivudine-resistant (LAM-R). The aim of this study was to compare the efficacy between TDF and TDF-based combination therapy against LAM-R HBV in CHB patients. Randomized and non-randomized control trials directly comparing TDF and TDF-based therapy for treatment of LAM-R CHB patients, were searched in Pubmed, Medline, EMBASE, database up to June 15, 2015. The data were analyzed with Review Manager (v.5.3). Five articles (683 patients in total) met entry criteria. The overall efficacy of tenofovir-based combination therapy was not significantly better with regard to the rates of virological response (85.5% vs. 81.5%; RR=0.95, 95%CI=0.88-1.03, P=0.25), ALT normalization (61.9% vs.72.0%; RR=1.18, 95%CI=0.96-1.44, P=0.11) and HBeAg loss (17.0% vs. 18.1%; RR=1.40, 95%CI=0.78-2.49, P=0.26) compared with TDF monotherapy through 48-week treatment. Additionally, subgroup analysis showed that no significant difference was determined as TDF group compared to TDF-based group at 48weeks, in terms of rates of HBV DNA undetectability, ALT normalization and HBeAg loss in the treatment of LAM-R patients with prior failure of LAM monotherapy. Moreover, the rates of HBV DNA suppression between groups were similar through 24 or 48weeks of treatment in LAM-R patients with prior failure of LAM/ADV therapy. TDF monotherapy is as effective as TDF-based combination therapy in maintaining viral suppression in LAM-R patients with prior failure of LAM or LAM/ADV therapy.


DeepStack-DTIs: Predicting Drug-Target Interactions Using LightGBM Feature Selection and Deep-Stacked Ensemble Classifier.
Accurate prediction of drug-target interactions (DTIs), which is often used in the fields of drug discovery and drug repositioning, is regarded a key challenge in the study of drug science. In this paper, a new method called DeepStack-DTIs is proposed to predict DTIs. First, for the target protein, pseudo-position specific score matrix, pseudo amino acid composition and SPIDER3 are used to extract the different feature information of the target protein. Meanwhile, the path-based fingerprint features of each drug are extracted. Then, the synthetic minority oversampling technique (SMOTE) and light gradient boosting machine (LightGBM) are used for data balancing and feature selection, respectively. Finally, the processed features are input to the deep-stacked ensemble classifier composed of gated recurrent unit (GRU), deep neural network (DNN), support vector machine (SVM), eXtreme gradient boosting (XGBoost) and logistic regression (LR) to predict DTIs. Under the five-fold cross-validation and compared with existing methods, the proposed method achieves higher prediction accuracy on the gold standard dataset. To evaluate the predictive power of DeepStack-DTIs, we validate the method on another dataset and predict the drug-target interaction network. The results indicate that DeepStack-DTIs has excellent predictive ability than the other methods, and provides novel insights for the prediction of DTIs. A novel method DeepStack-DTIs for drug-target interactions prediction. PsePSSM, PseAAC, SPIDER3 and FP2 are fused to convert protein sequence and drug molecule information into digital information, respectively. The SMOTE algorithm is used to balance the dataset and LightGBM feature selection algorithm is employed to remove redundant and irrelevant features to select the optimal feature subset. This optimal feature subset is inputted into the deep-stacked ensemble classifier to predict drug-target interactions. The experimental results show DeepStack-DTIs method can significantly improve the prediction accuracy of drug-target interactions.


Local immunomodulation with CD4 and CD8 antibodies, but not cyclosporine A, improves osteogenesis induced by ADhBMP9 gene therapy.
This study was designed to see if immunosuppression achieved using local application of cyclosporine A (Cs. A) or CD4 and CD8 antibodies would improve bone formation following intramuscular injections of human BMP-4 and BMP-9 adenoviral vectors (ADhBMP4 and ADhBMP9) in Sprague-Dawley rats. Cs. A was injected into the thigh muscle. After 2 days, ADhBMP4, ADhBMP9, and the antibodies were separately injected into the left and right rear legs. At this time, the number of CD4+/CD3+ cells was significantly lower and the number of CD8+/CD3+ cells higher in the Cs. A group than in the control group (P < 0.01). The total number of white blood cells 3 days following injection of CD4 and CD8 antibodies was significantly lower than that before the injection (P < 0.01). At 4 weeks after the viral and antibody injections, mean bone volumes at the ADhBMP9 treatment sites were 0.29 +/- 0.01 cm3 in the viral control group, 0.17 +/- 0.03 cm3 in the Cs. A-ADhBMPs group, and 0.59 +/- 0.07 cm3 in the antibodies-ADhBMPs group. ADhBMP4 did not induce new bone formation in any group. This study demonstrates that local immunomodulation may improve the osteogenic potential of bone morphogenetic protein gene therapy in the clinical setting.


2-(5-Fluoro-2,3-dioxoindolin-1-yl)ethyl 4-methyl-piperazine-1-carbodithio-ate.
In the title compound, C(16)H(18)FN(3)O(2)S(2), the methyl-piperazine ring adopts a chair conformation, while the (2,3-dioxoindolin-1-yl)ethyl unit is linked to one of the N atoms of the piperazine ring via the carbodithio-ate group. In the crystal, each mol-ecule is linked to its neighbors within the ([Formula: see text]03) plane through weak C-H(methyl-ene)⋯O, C-H(ar-yl)⋯O and C-H(methyl-ene)⋯S inter-actions. Perpendicular to this plane mol-ecules are connected through inter-molecular short N⋯π(pyrrole ring) contacts [N⋯C centroid = 3.232 (2) Å], another set of C-H(methyl-ene)⋯O inter-actions and through short contacts between carbodithio-ate S atoms and the pyrrole rings [C⋯centroid = 3.695 (3), S⋯centroid = 3.403 (2) Å].


Animal models of spontaneous and drug-induced cutaneous lupus erythematosus.
Skin lesions are one of the most common manifestations of lupus erythematosus (LE) disorders such as systemic LE and discoid LE. The etiology of cutaneous LE is not fully understood. To address this issue, appropriate animal models frequently clarify the etiology and pathogenesis of autoimmune diseases, although no single animal model perfectly mimics a human disease. A common dermatological finding in many SLE-prone mouse strains is the deposition of immunoglobulins at the dermoepidermal junction. Over the past decade, the most exciting and important finding has been the discovery of the Fas-defect in the pathogenesis of the autoimmune MRL/lpr mouse, which is a good model for the spontaneous development of skin lesions similar to those seen in human LE. The analysis of MRL/lpr mice showed a close association between immunoglobulin deposits and the appearance of skin lesions. Transgenic and knock out mice have advanced the investigation of cutaneous LE. Furthermore, the model of drug-induced cutaneous LE can yield additional insight since the trigger is clear in drug-induced LE. Cutaneous LE lesions can also be induced in TCRalpha-/- mice treated with fluorouracil and ultraviolet B light irradiation. Studies on both spontaneous and experimental models will elucidate the pathogenesis of complicated and multifactorial cutaneous LE.


Two novel mutations and a previously unreported intronic polymorphism in the NOTCH3 gene.
Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a hereditary disease of small vessel caused by mutations in the NOTCH3 gene (NCBI Gene ID: 4854) located on chromosome 19p13.1. NOTCH3 consists of 33 exons which encode a protein of 2321 amino acids. Exons 3 and 4 were found to be mutation hotspots, containing more than 65% of all CADASIL mutations. We performed direct sequencing on an ABI 3130 Genetic Analyser to screen for mutations and polymorphisms on 300 patients who were clinically suspected to have CADASIL. First, exons 3 and 4 were screened in NOTCH3 and if there were no variations found, then extended CADASIL testing (exons 2, 11, 18 and 19) was offered to patients. Here we report two novel non-synonymous mutations identified in the NOTCH3 gene. The first mutation, located in exon 4 was found in a 49-year-old female and causes an alanine to valine amino acid change at position 202 (605C>T). The second mutation, located in exon 11, was found in a 66-year-old female and causes a cysteine to arginine amino acid change at position 579 (1735T>C). We also report a 46-year-old male with a known polymorphism Thr101Thr (rs3815188) and an unreported polymorphism NM_000435.2:c.679+60G>A observed in intron 4 of the NOTCH3 gene. Although Ala202Ala (rs1043994) is a common polymorphism in the NOTCH3 gene, our reported novel mutation (Ala202Val) causes an amino acid change at the same locus. Our other reported mutation (Cys579Arg) correlates well with other known mutations in NOTCH3, as the majority of the CADASIL-associated mutations in NOTCH3 generally occur in the EGF-like (epidermal growth factor-like) repeat domain, causing a change in the number of cysteine residues. The intronic polymorphism NM_000435.2:c.679+60G>A lies close to the intron-exon boundary and may affect the splicing mechanism in the NOTCH3 gene.


Identification of circ_0038632 as a Promoter of Breast Cancer through miR-520a-3p-Dependent Modulation of CDCA3.
Emerging evidence proves the importance of circular RNAs (circRNAs) in many tumors, including breast cancer (BC). Here, we aimed to define a mechanism by which circ_0038632 regulates BC process. To quantify the expression of circ_0038632, miR-520a-3p and cell division cycle associated 3 (CDCA3), a quantitative real-time PCR or immunoblotting method was utilized. The relationships of circ_0038632/miR-520a-3p and miR-520a-3p/CDCA3 in BC cells were determined using RNA immunoprecipitation (RIP) experiment and luciferase reporter assay. The effects of the circ_0038632/miR-520a-3p/CDCA3 cascade on cell biological phenotypes in vitro were examined by flow cytometry, EdU assay, cell counting kit 8 assay, transwell assay and wound healing assay. The function of circ_0038632 in tumorigenicity of BC cells in vivo was evaluated by xenograft experiments. Circ_0038632 and CDCA3 were highly expressed and miR-520a-3p expression was hindered in human BC. Depletion of circ_0038632 weakened cell growth, motility, and invasiveness while promoted cell apoptosis. In terms of mechanism, miR-520a-3p targeted CDCA3, and circ_0038632 involved the post-transcriptional modulation of CDCA3 expression by working as a miR-520a-3p sponge. Silencing of miR-520a-3p could reverse the inhibitory functions of circ_0038632 depletion in BC cell malignant phenotypes. Re-expression of CDCA3 also overturned the suppressive effects of miR-520a-3p on BC cell malignant phenotypes. In addition, circ_0038632 depletion inhibited the growth of xenograft tumors in vivo. Taken together, circ_0038632 promotes breast carcinogenesis through the miR-520a-3p/CDCA3 regulatory cascade, indicating that circ_0038632 may be a potential target for BC treatment.


Regional variation in blindness in children due to microphthalmos, anophthalmos and coloboma.
The prevalence and causes of blindness in children vary widely between regions. Few epidemiological data are available on the relative importance of the major congenital anomalies of the globe (i.e., microphthalmos, anophthalmos, coloboma) as causes of blindness in children. The aim of this study was to determine the re-gional variation in the proportion of severe visual impairment and blindness due to congenital abnormalities of the globe in children in schools for the blind and in those identified through Community Based Rehabilitation programs. Other objectives were to estimate the prevalence of blindness due to major congenital abnormalities, and to investigate their etiology. Data on the causes of blindness in children were collected between 1990 and 1998 using standard methods, definitions and reporting form in 26 countries. Children were examined in schools for the blind and in Community Based Rehabilitation programs. Of 7,113 children aged 3-15 years with severe visual impairment and blindness examined, 762 (10.7%) had microphthalmos, 161 (2.3%) had anophthalmos, and 96 (1.3%) had coloboma. There are large regional differences in the proportion of severe visual loss in blind school children, ranging from 1.4% in Cuba to 33.2% in Sri Lanka. Severe visual loss due to congenital abnormalities of the globe is estimated to affect between 0.4 and 16.2/100,000 children in the countries studied. An underlying cause could not be identified in 84.2%. Major congenital abnormalities of the globe are important causes of severe visual loss in children, particularly in Asian countries. Further research into etiology is warranted in order to plan prevention programs.


An improved method for preparing stained ground teeth sections.
In oral histopathology teaching and research, there is a need for high-quality undemineralized tooth sections that are easy to handle, have controlled thickness, allow the observation of intact microstructures, and can be preserved for long periods of time. Teeth were collected under non-demineralizing conditions. Tooth sections (15-25 µm) were prepared using a diamond knife, then randomly divided into three groups: (1) stained with rosin, (2) stained with hematoxylin and eosin, or (3) not stained. The prepared tooth sections were evaluated by microscopy for clarity and microstructure visibility. The use of a diamond knife in the sectioning and grinding process yielded high-quality ground sections of teeth. Rosin-stained ground sections allowed better identification of microstructures within the teeth, compared with unstained or hematoxylin and eosin-stained ground sections. The best results were obtained in the ground sections of teeth that were stained with rosin. Ground sections of teeth prepared using this staining method could be useful in oral histopathology teaching and research.


The toxin-agglutinin fold. A new group of small protein structures organized around a four-disulfide core.
The three-dimensional structures of the snake venom postsynaptic neurotoxins and of the domains in wheat germ agglutinin show a remarkably similar overall folding pattern, consisting of equivalently placed, but variably sized loops which are held together by four similarly positioned disulfide bonds. Furthermore, occurrence of this wheat germ agglutinin-neurotoxin domain fold is predicted not only in the snake venom cardiotoxins and cytotoxins with neurotoxin-matched half-cystine sequence positions, but also for two small plant proteins, hevein and ragweed pollen allergen Ra5, on the basis of a nearly exact match of their half-cystine, sequence positions with those of the wheat germ agglutinin domain.


Mouse model of proximal colon-specific tumorigenesis driven by microsatellite instability-induced Cre-mediated inactivation of Apc and activation of Kras.
KRAS gene mutations are found in 40-50% of colorectal cancer cases, but their functional contribution is not fully understood. To address this issue, we generated genetically engineered mice with colon tumors expressing an oncogenic Kras(G12D) allele in the context of the Adenomatous polyposis coli (Apc) deficiency to compare them to tumors harboring Apc deficiency alone. CDX2P9.5-G22Cre (referred to as G22Cre) mice showing inducible Cre recombinase transgene expression in the proximal colon controlled under the CDX2 gene promoter were intercrossed with Apc (flox/flox) mice and LSL-Kras (G12D) mice carrying loxP-flanked Apc and Lox-Stop-Lox oncogenic Kras(G12D) alleles, respectively, to generate G22Cre; Apc(flox/flox); Kras(G12D) and G22Cre; Apc(flox/flox); KrasWT mice. Gene expression profiles of the tumors were analyzed using high-density oligonucleotide arrays. Morphologically, minimal difference in proximal colon tumor was observed between the two mouse models. Consistent with previous findings in vitro, Glut1 transcript and protein expression was up-regulated in the tumors of G22Cre;Apc (flox/flox) ; Kras(G12D) mice. Immunohistochemical staining analysis revealed that GLUT1 protein expression correlated with KRAS mutations in human colorectal cancer. Microarray analysis identified 11 candidate genes upregulated more than fivefold and quantitative PCR analysis confirmed that Aqp8, Ttr, Qpct, and Slc26a3 genes were upregulated 3.7- to 30.2-fold in tumors with mutant Kras. These results demonstrated the validity of the G22Cre; Apc(flox/flox) ;Kras (G12D) mice as a new mouse model with oncogenic Kras activation. We believe that this model can facilitate efforts to define novel factors that contribute to the pathogenesis of human colorectal cancer with KRAS mutations.


Pneumocephalus secondary to spontaneous bronchial-subarachnoid fistula.
We describe a patient in whom pneumocephalus appeared spontaneously, resulting from a communication between the bronchial tree and the thoracic subarachnoid space in association with squamous cell carcinoma of the lung. The communication was demonstrated by myelography and obliterated surgically. A review of the literature is included.


Effects of 2-methoxyethanol on fetal development, postnatal behavior, and embryonic intracellular pH of rats.
The industrial solvent 2-methoxyethanol (2ME) is a reproductive and developmental toxicant when administered by inhalation, gavage, and IP injection. The present research established that this solvent can produce teratogenicity in rats when administered in liquid diet. Groups of 10 Sprague-Dawley rats were given various percentages of 2ME in liquid diet on gestation days 7-18. Day 20 fetuses were examined for visceral or skeletal malformations. Concentrations above 0.025% 2ME (approximately 73 mg/kg/day) produced total embryo-mortality. Cardiovascular malformations were produced at lower levels. The teratogenic no-effect level was 0.006% 2ME (16 mg/kg). In a second experiment, groups of 12 Sprague-Dawley rats were given 0, 0.006 and 0.012% of 2ME as above. Litters were culled to 8 pups, and tested for auditory and tactile startle and conditioned lick suppression, and for performance in figure-8 activity and the Cincinnati water maze on postnatal days 48-65. The high dose of 2ME produced approximately 50% mortality in the offspring and increased the number of errors in the Cincinnati maze. No other behavioral effects were observed at either dose. An interaction study was conducted to determine if simultaneous exposure to 2ME and ethanol would reduce the teratogenicity of 2ME, but no reduction was observed. The hypothesis that 2ME acts by altering embryonic intracellular pH was tested by injecting 0.33 ml/kg of 2ME into rats on gestation day 13, and determining embryonic intracellular pH at 2, 4, 8, and 24 hours thereafter. There was an increase in pH at 4 hours, but not at later time points. Another group of rats was given 2ME along with amiloride, which blocks the sodium/hydrogen antiporter. The combined 2ME-amiloride exposure produced an incidence of cardiovascular malformations in fetuses twice that of 2ME alone. These studies confirmed the structural teratogenicity of 2ME even when given in liquid diet, as it was given for the first time in the present study. At nonteratogenic doses, developmental toxicity (e.g., postnatal deaths) persisted, but only limited evidence of behavioral teratogenicity was observed. The pH data are consistent with the concept that 2ME may alter embryonic intracellular pH at critical stages of organogenesis.


Limits on Anti-Phase Synchronization in Oscillator Networks.
Anti-phase synchronization is the spontaneous formation of 2 clusters of oscillators synchronized between themselves within a cluster but opposite in phase with the other cluster. Neuronal networks in human and animal brains, ecological networks, climactic networks, and lasers are all systems that exhibit anti-phase synchronization although the phenomenon is encountered less frequently than the celebrated in-phase synchronization. We show that this disparity in occurrence is due to fundamental limits on the size of networks that can sustain anti-phase synchronization. We study the influence of network structure and coupling conditions on anti-phase synchronization in networks composed of coupled Stuart-Landau oscillators. The dependence of probability of anti-phase synchronization on connectivity of the network, strength of interaction over distance, and symmetry of the network is illustrated. Regardless of favourable network conditions, we show that anti-phase synchronization is limited to small networks, typically smaller than 20 nodes.


Vertebral artery injury in patients with isolated transverse process fractures.
We sought to assess the rate of CTA-diagnosed vertebral artery injury in patients with isolated transverse process fractures, with and without extension into the transverse foramen, in the blunt-trauma population served by our hospital. We queried our universities trauma registry between January 2009 and July 2014 for ICD-9 codes pertaining to cervical spine fractures. Of 330 patients identified, 45 patients had fractures limited to the transverse process and were selected for the study population. For each patient identified, demographics, injury mechanism, imaging reports, angiography findings, and treatments were recorded. In total, 69 fractures were identified in 45 patients. Of the 45 patients, 15 (33%) had transverse process fractures at multiple cervical levels. 23/45 (51%) patients had at least one fracture extending into TF. Four patients with transverse process fractures and one patient without transverse process fractures were diagnosed with vertebral artery injury by CT angiogram (17.4% vs. 4.5%, p=0.35). The number of transverse process fractures in patients with VAI was greater than those without VAI (3.0 vs. 1.4, p<0.001). None of the 30 patients with any one-level TPF (with or without extension into TF) was diagnosed with VAI (p=0.003). None of 17 patients with isolated C7-level TPFs were diagnosed with VAI (p=0.15). The incidence of cervical VAI was greater in patients with multiple-level TPFs than in patients with single-level TPFs. While patients with a single, isolated TPF have a low probability of VAI, patients with numerous TPF fractures may benefit from CTA.