miR-429 inhibits migration and invasion of breast cancer cells in vitro.
Accumulating evidence indicates that microRNAs (miRNAs) are involved in regulating cancer invasion and metastasis, and an increasing number of research demonstrates that miRNAs can promote or inhibit cell motility depending on genetic background of different cancers and the microenvironment. In the present study, we established an in vivo bone metastasis model of breast cancer by injecting MDA-MB-231 cells into the left ventricle of nude mice, and then screened the differentially expressed miRNAs between parental and bone-metastatic MDA-MB-231 cells using miRNA array. The results revealed that decreased expression of miR-429 was probably involved in negatively regulating bone metastasis of breast cancer cells. On the other hand, overexpression of miR-429 in MDA-MB-231 cells remarkably suppressed invasion in vitro. We identified ZEB1 and CRKL as potential targets of miR-429 by analyzing combined results from in silico search and global expression array of the same RNA samples. Immunoblot assay confirmed that miR-429 reduced their expression at protein level. Taken together, our results offer an opportunity for further understanding of the recondite mechanisms underlying the bone metastasis of breast cancer.


A highly reproducible method of nucleolus organizing regions staining in plants.
A method for the specific detection of the nucleolus organizing regions (NORs) of plant chromosomes has been developed employing enzymatic maceration and successive flame-drying for chromosome spreading and incubation with aqueous 50% AgNO3 at 55-60 C. When this method was applied to metaphase chromosomes the NORs were specifically discriminated as heavily stained segments in all the plant species examined. In the satisfactory results obtained by monitoring the reaction under a microscope during the course of the silver treatment, the chromosome arms were stained yellow to light brown while the NORs were dark brown to black. The present method has the advantage of yielding highly reproducible results for the specific detection of the NORs in plant materials.


Histamine skin reactivity increases with body mass index in Korean children.
Histamine skin prick testing is most commonly used to diagnose immunoglobulin E (IgE)-mediated allergic diseases, and histamine reactivity is used as a standardized positive control in the interpretation of a skin prick test. However, reactivity to histamine differs among individuals for reasons that are poorly understood. The present study aimed to evaluate the potential association between body mass index (BMI) and histamine skin reactivity in children. A total of 451 children (246 boys, 205 girls) aged 7-8 years were enrolled in this study. The skin prick test was performed with 26 aeroallergens commonly found in Korea. Other information was collected, including sex, age, BMI, parental allergy history, and parental smoking status. Multivariate analysis was used to confirm the association between histamine skin reactivity and BMI. The histamine wheal size was revealed to be associated with BMI (Spearman's Rho 0.161, p<0.001). This association was confirmed by multivariate analysis, after adjusting for sex, age, parental allergy history, parental smoking status, and allergic sensitization (coefficient B 0.071, 95% confidence interval 0.030-0.112). Skin responses to histamine were primarily correlated with increased BMI. Further studies are needed to understand the clinical implication of BMI when interpreting the results of skin prick test.


Congenital toxoplasmic retinochoroiditis in a mouse model.
A study of the eyes of adult mice infected in utero with Toxoplasma gondii is reported. The histopathological features of the ocular inflammatory response in the infected mice ranged from minimal damage to complete destruction of the retinal tissue. Notable features such as retinal vasculitis and an almost uniform and highly selective destruction of the photoreceptor layer of the retina suggest a similarity between experimental autoimmune retinitis and the disease process in the retinas of our Toxoplasma-infected mice. We suggest that our mouse model could provide a simple and inexpensive tool for the investigation of immuno-pathological processes in the retina resulting from congenital Toxoplasma infection. The model has the advantage of low post-natal mortality coupled with high ocular morbidity. Furthermore, its aetiology is probably analogous to that of human ocular toxoplasmosis, in that the foetus becomes infected in utero via a mother whose primary infection is acquired during gestation.


Magnetic resonance-guided focused ultrasound myomectomy: safety, efficacy, subsequent fertility and quality-of-life improvements, a systematic review.
We performed a systematic review about studies reporting data of myomectomy performed by magnetic resonance-guided focused ultrasound (MRgFUS) technique in order to define its safety, feasibility, indications, complications, and impact on uterine fibroid symptom and health-related quality of life (UFS-QOL) and fertility. Outcomes were considered according to fibroids shrinkage, nonperfused volume (NPV), NPV ratio, and uterine fibroid symptoms assessed with UFS-QOL questionnaire (baseline 3, 4, 6, and 12 months). We analyzed 38 eligible studies reporting outcomes about 2500 patients (mean age 43.67 years). The MRgFUS results a safe, efficient, and cost-effective minimal invasive technique for treatment of uterine fibroids. Increasing experience, device improvements, and availability for a larger number of patients are enhancing the outcomes, while the obstetrical ones should be more extensively explored. The MRgFUS could be considered as a minimal invasive alternative to traditional surgical or radiological procedures for the treatment of symptomatic uterine myomas improving both QOL and subsequent fertility.


Hemodynamic changes before acute heart failure episodes in patients with advanced systolic left ventricular dysfunction.
We sought to verify which changes in right ventricular pressures occur before acute heart failure (HF) in patients with advanced systolic left ventricular (LV) dysfunction. In patients with chronic HF, continuous hemodynamic monitoring by implantable devices may detect increases in pulmonary pressures before the onset of symptoms. A device which allowed monitoring of right ventricular systolic (RVSP) and diastolic pressure (RVDP) and an estimate of pulmonary artery diastolic pressure (ePAD) was implanted in 10 patients with advanced LV systolic dysfunction and frequent cardiovascular hospitalizations [mean age 56.8 years; New York Heart Association (NYHA) classes IIIb-IV; LV ejection fraction at echocardiography 21 +/- 2%]. During a follow-up period of 15 +/- 12 months, 18 hospitalizations due to acute HF were recorded; 10 episodes were characterized by symptoms due to low-cardiac output (LCO) and eight episodes were characterized by symptoms due to pulmonary congestion. RVSP and ePAD increased before six hospitalizations and decreased before three episodes; RVDP increased before 10 hospitalizations and decreased before one. The extent of RVDP increase was 16 +/- 24% before pulmonary congestion episodes and 29 +/- 32% before LCO episodes. An increase in RVDP was the most frequent hemodynamic change detected by the implantable hemodynamic monitor before hospitalizations due to acute HF in patients having advanced systolic LV dysfunction.


The automated physiologic profile.
The automated physiologic profile provides the critical care physician with hemodynamic, oxygen consumption and tissue utilization data at reasonable cost. A paramedical assistant performs all data acquisition, recordings and blood sampling procedures. Data reduction is performed through use of off-the-shelf desk-top calculator equipment and accessories and a standardized graphic display is provided for the physician in charge. The physiologic profile has been utilized in high-risk patients requiring medical and surgical interventions. It is routinely employed in postoperative monitoring of cardiac patients and patients undergoing major surgical procedures with associated cardiovascular decompensation. Diagnosis of cardiac and pulmonary deterioration and promptness of advanced support interventions represent additional areas of effective clinical application.


On the mutagenicity and immunogenicity of a beta-propiolactone-treated therapeutic IgG-preparation.
During the production of Intraglobin, an i.v. tolerable IgG-immunoglobulin preparation, fraction Cohn II is treated with beta-propiolactone. Thereby the IgG-molecules are chemically modified thus preventing an increase in secondary aggregates which could give rise to adverse reactions in the patients treated with the IgG-preparation. The safety tests described here neither showed any evidence for the presence of unhydrolysed mutagenic-beta-propiolactone in the final product nor that neoantigenic determinants had been created by the reaction of beta-propiolactone with the protein molecules.


Prognosis of direct pregnancy in untreated atypical endometrial hyperplasia: a case report.
An increasing number of atypical endometrial hyperplasia (AEH) or endometrial cancer (EC) patients with fertility requirements choose conservative management, such as oral high-dose progesterone. Most of them use assisted reproductive technology (ART) to become pregnant after experiencing remission. However, the outcome of pregnancy is not ideal, probably because of long-term drug application in large doses or invasive uterine cavity treatment. We presented a case of AEH who underwent direct pregnancy with good results without any treatment for her pathological endometrium. We described her endometrial histological results pre-and post-pregnancy in detail, hitherto absent from reports on this topic. Patients with a strong desire to bear children at the time of an AEH diagnosis could consider taking 1-2 years to try a pregnancy before treating their AEH.


Management of complications after skin surgery relevant for melanoma in the trunk and extremities during the COVID-19 pandemic: a case series report.
Patients with melanoma have been found to be at greater risk of adverse outcomes including mortality after contacting COVID-19. Management of postsurgical complications presented additional challenges by potentially increasing exposure to COVID-19 through repeated inpatient admissions to hospital during the pandemic. We report four cases for which skin flaps, lymph ligation, and split-thickness skin graft (STSG) were successfully used in the treatment of complications in the trunk and extremities after wide local excision (WLE). This study details the operative experience in management of postsurgical complications for melanoma in the trunk and extremities during a 6-month period at the height of the COVID-19 pandemic. We present 4 cases detailing management of complications that occurred after wide local excisions performed for melanoma during Feb. to Oct. 2020. Case 1: A 90-year-old man who experienced wound dehiscence and necrosis on the shoulder after non-radical excision for an aggressive melanoma and underwent the side-to-side closure after ellipse formed WLE with modified tangent-to-circle method. Case 2: An 80-year-old man who had undergone excision for melanoma in his left upper arm and histopathology did not show radically. Two weeks after the excision, he underwent a WLE and direct reconstruction with double rotation skin flap. Case 3: A 55-year-old man that experienced a large wound dehiscence on his back due to WLE. He underwent an advanced double skin flap operation. Case 4: A 36-year-old woman who had a lymphorrhea and graft necrosis after WLE and STSG on the right lower leg. A combination of micro lymph ligation and re-STSG was performed. One month after the operation, all wounds had healed. There was no clinical evidence of tumor recurrence after 8 months post procedure. Severe complications (e.g., large wound dehiscence, necrosis, or lymphorrhea) following wide local excision of melanoma are infrequent but must be swiftly and appropriately managed, especially during the COVID-19 pandemic to decrease the likelihood of COVID-19 infection and impaired oncology outcomes from delaying systemic cancer therapy due to the complications in primary interventions.


A Machine Learning Workflow of Multiplexed Immunofluorescence Images to Interrogate Activator and Tolerogenic Profiles of Conventional Type 1 Dendritic Cells Infiltrating Melanomas of Disease-Free and Metastatic Patients.
Melanoma is the deadliest form of skin cancer. Due to its high mutation rates, melanoma is a convenient model to study antitumor immune responses. Dendritic cells (DCs) play a key role in activating cytotoxic CD8+ T lymphocytes and directing them to kill tumor cells. Although there is evidence that DCs infiltrate melanomas, information about the profile of these cells, their activity states, and potential antitumor function remains unclear, particularly for conventional DCs type 1 (cDC1). Approaches to profiling tumor-infiltrating DCs are hindered by their diversity and the high number of signals that can affect their state of activation. Multiplexed immunofluorescence (mIF) allows the simultaneous analysis of multiple markers, but image-based analysis is time-consuming and often inconsistent among analysts. In this work, we evaluated several machine learning (ML) algorithms and established a workflow of nine-parameter image analysis that allowed us to study cDC1s in a reproducible and accessible manner. Using this workflow, we compared melanoma samples between disease-free and metastatic patients at diagnosis. We observed that cDC1s are more abundant in the tumor infiltrate of the former. Furthermore, cDC1s in disease-free patients exhibit an expression profile more congruent with an activator function: CD40highPD-L1low CD86+IL-12+. Although disease-free patients were also enriched with CD40-PD-L1+ cDC1s, these cells were also more compatible with an activator phenotype. The opposite was true for metastatic patients at diagnosis who were enriched for cDC1s with a more tolerogenic phenotype (CD40lowPD-L1highCD86-IL-12-IDO+). ML-based workflows like the one developed here can be used to analyze complex phenotypes of other immune cells and can be brought to laboratories with standard expertise and computer capacity.


Mapping of nascent light and heavy strand transcripts on the physical map of HeLa cell mitochondrial DNA.
The sequences complementary to the nascent RNA molecules isolated from transcription complexes of HeLa cell mtDNA have been mapped on the H and L strands of mtDNA by the S1 protection technique. The distribution of these sequences among different Hpa II restriction fragments was found to reflect the position of these fragments in the Hpa II map of mtDNA. Thus, the S1-resistant hybrids formed with the L strand corresponded almost exclusively to the right half of the genome past the origin of replication in the direction of L strand transcription, and were especially concentrated in the region immediately adjacent to the origin. By contrast, the hybrid duplexes involving the H strand appeared to be localized in the left half of the genome, and in particular in the quadrant of the map adjacent to the origin in the direction of H strand transcription. These results strongly suggest that the region of mtDNA around the origin of replication contains an initiation site for L strand transcription and an initiation site for H strand transcription.


Thermal Conductivity and Mechanical Properties of Thermoplastic Polyurethane-/Silane-Modified Al2O3 Composite Fabricated via Melt Compounding.
The increase of miniaturization and rise of powerhouses has caused a need for high-performing thermal interface materials (TIMs) that can transfer heat in electronic packaging. In this study, a thermoplastic polyurethane (PU)/alumina composite was produced via twin extrusion and was suggested as a TIM. The surfaces of the alumina particles were modified by γ-aminopropyltriethoxysilane (APTES) and then evaluated using Fourier transform infrared spectroscopy (FT-IR) and X-ray photoelectron spectroscopy (XPS). The field emission scanning electron microscopy (FE-SEM) images revealed that the addition of surface-modified alumina was well adhered in the PU matrix. The tensile strength of the composite remained unchanged, while the Young's modulus showed improvement as compared to the pure PU. The elongation at the break decreased as the filler loading increased, due to the brittle behavior of the composite. The viscoelastic elastic property analysis results revealed that there was an increase in the storage modulus of the composite and the glass transition temperature curve shifted to the right. The thermal conductivity of the composite showed that there was an 80.6% improvement in thermal conductivity with the incorporation of 40% APTES-treated alumina particles.


Bibliometric analysis of RNA vaccines for cancer.
Cancer represents a challenging medical problem worldwide. Several exploratory studies have been conducted to overcome these limitations. RNA vaccines play an important role in cancer prevention and treatment. Recent studies have shown that RNA vaccines play an important role in cancer prevention. However, there are currently no relevant bibliometric studies. This study aimed to apply bibliometrics to summarize the knowledge structure and research hotspots regarding the role of RNA vaccines in cancer. Publications related to RNA vaccines in cancer were searched on the web of science core collection (WoSCC) database. VOSviewers, CiteSpace and R package "bibliometrix" were used to conduct this bibliometric analysis. A total of 1399 articles were included, comprising 803 original articles and 596 reviews. The number of studies on RNA vaccines against cancer has been increasing annually. China and the United States were the principal countries of origin of publications. Johannes Gutenberg University Mainz, NCI, and Duke University were the main organizations. Frontiers in Immunology is the leading journal in the field. Hot keywords included tumor antigens, lipid nanoparticles, emerging roles, and dendritic cells. This is the bibliometric study to summarize the research trends and development of RNA vaccines for cancer. This information will provide a reference for researchers studying RNA vaccines against cancer.


Autophagy: more than a nonselective pathway.
Autophagy is a catabolic pathway conserved among eukaryotes that allows cells to rapidly eliminate large unwanted structures such as aberrant protein aggregates, superfluous or damaged organelles, and invading pathogens. The hallmark of this transport pathway is the sequestration of the cargoes that have to be degraded in the lysosomes by double-membrane vesicles called autophagosomes. The key actors mediating the biogenesis of these carriers are the autophagy-related genes (ATGs). For a long time, it was assumed that autophagy is a bulk process. Recent studies, however, have highlighted the capacity of this pathway to exclusively eliminate specific structures and thus better fulfil the catabolic necessities of the cell. We are just starting to unveil the regulation and mechanism of these selective types of autophagy, but what it is already clearly emerging is that structures targeted to destruction are accurately enwrapped by autophagosomes through the action of specific receptors and adaptors. In this paper, we will briefly discuss the impact that the selective types of autophagy have had on our understanding of autophagy.


Low-Volume Bowel Preparation Is Associated With Reduced Time to Colonoscopy in Hospitalized Patients: A Propensity-Matched Analysis.
Delays in inpatient colonoscopy are commonly caused by inadequate bowel preparation and result in increased hospital length of stay (LOS) and healthcare costs. Low-volume bowel preparation (LV-BP; sodium sulfate, potassium sulfate, and magnesium sulfate ) has been shown to improve outpatient bowel preparation quality compared with standard high-volume bowel preparations (HV-BP; polyethylene glycol ). However, its efficacy in hospitalized patients has not been well-studied. We assessed the impact of LV-BP on time to colonoscopy, hospital LOS, and bowel preparation quality among inpatients. We performed a propensity score-matched analysis of adult inpatients undergoing colonoscopy who received either LV-BP or HV-BP before colonoscopy at a quaternary academic medical center. Multivariate regression models with feature selection were developed to assess the association between LV-BP and study outcomes. Among 1,807 inpatients included in this study, 293 and 1,514 patients received LV-BP and HV-BP, respectively. Among the propensity score-matched population, LV-BP was associated with a shorter time to colonoscopy (β: -0.43 [95% confidence interval: -0.56 to -0.30]) while having similar odds of adequate preparation (odds ratio: 1.02 [95% confidence interval: 0.71-1.46]; P = 0.92). LV-BP was also significantly associated with decreased hospital LOS among older patients (age ≥ 75 years), patients with chronic kidney disease, and patients who were hospitalized with gastrointestinal bleeding. LV-BP is associated with decreased time to colonoscopy in hospitalized patients. Older inpatients, inpatients with chronic kidney disease, and inpatients with gastrointestinal bleeding may particularly benefit from LV-BP. Prospective studies are needed to further establish the role of LV-BP for inpatient colonoscopies.


[The health of unemployed women and men. First results of the Telephone Health Survey 2003].
Unemployment has material and psychosocial consequences that can also have an adverse effect on health. According to the Telephone Health Survey 2003, unemployed women and men suffer more frequently from many diseases and complaints. Their assessment of their own state of health is also more negative, and they say more often that their quality of life is restricted by their health. In the case of men, health problems are especially common among the long-term unemployed (>1 year), whereas women can suffer equally (or even more) serious effects on their health as a result of short-term unemployment. On the other hand, the analysis clearly indicates that poor health heightens a person's risk of becoming unemployed. However, no conclusive statement can be made on the direction of causality between unemployment and health, since this requires more longitudinal data.


Seasonality, Composition, and Antioxidant Capacity of Limonene/δ-3-Carene/(E)-Caryophyllene Schinus terebinthifolia Essential Oil Chemotype from the Brazilian Amazon: A Chemometric Approach.
Schinus terebinthifolia Raddi is widely used in traditional Brazilian medicine to treat respiratory diseases, as an antiseptic, anti-inflammatory, and hemostatic agent. This study aimed to evaluate the influence of climatic parameters on the yield, antioxidative capacity, and chemical composition of the S. terebinthifolia leaf essential oil. The specimen was collected monthly from October 2021 to September 2022. Leaf essential oils (EOs) were obtained by hydrodistillation, and their chemical compositions were analyzed by gas chromatography/mass spectrometry (GC/MS). Statistical analyses were performed to verify the climatic influences on the yields, chemical composition, and antioxidative capacity. The DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging and inhibition of β-carotene/linoleic acid oxidation assays were performed to assess the antioxidant activity. The leaf essential oil yields ranged from 0.1% (July) to 0.7% (May and September), averaging 0.5 ± 0.2%. There was no significant difference in essential oil production during the dry (0.4 ± 0.2%) and rainy (0.6 ± 0.1%) seasons. The main chemical constituents identified in essential oils were limonene (11.42-56.24%), δ-3-carene (8.70-33.16%) and (E)-caryophyllene (4.10-24.98%). The limonene annual average was 43.57 ± 12.74% and showed no statistical difference during the dry (40.53 ± 13.38%) and rainy (52.68 ± 3.27%) seasons. Likewise, the annual average of δ-3-carene was 22.55 ± 7.11%, displaying no statistical difference between dry (26.35 ± 7.90%) and rainy (31.14 ± 1.63%) seasons. The annual average of (E)-caryophyllene was 11.07 ± 7.15% and this constituent did not show a statistical difference in Tukey's test (p > 0.05) during the dry (12.72 ± 7.56%) and rainy (6.10 ± 1.78%) season. Limonene showed a moderate positive and significant correlation (p < 0.05) with precipitation (r = 0.56) and a weak correlation with temperature (r = -0.40), humidity (r = 0.40), and insolation (r = -0.44). All samples inhibited the oxidation in the β-carotene/linoleic acid system (22.78-44.15%) but displayed no activity in the DPPH method.


Single-Center Experience in Vaccination of Children in Special Risk Groups: A Multidisciplinary Institutional Consensus Protocol.
Despite marked improvements in the accessibility of childhood vaccines, knowledge gaps remain about the vaccination of children in special risk groups (SRG). This study aimed to analyze the clinical data of children vaccinated in SRG in a single-center unit to contribute to the clinical evidence for the specific planning of immunization of children in SRG. The second- ary aim is to present institutional consensus on the vaccination of children in SRG. This retrospective study was conducted at a single-center pediatric vaccination clinic. Patient charts between 2018 and 2021 were retrospectively reviewed, and clinical and laboratory data were extracted. Serial joint meetings with multiple healthcare pro- fessionals were performed to develop an institutional protocol for vaccination. There were 479 children vaccinated between 2018 and 2021 for reasons such as post- chemotherapy, after hematopoietic stem cell transplantation, before/after solid organ trans- plantation, allergies, and chronic diseases. Of these, 298 (62.2%) children vaccinated in the unit due to a history of food or vaccine allergies were excluded. One hundred eighty-one children were vaccinated at a median age of 11 [7-15] years. Most children were vaccinated after treat- ment for malignancies. Solid tumors were the most frequent malignancy (67%), followed by acute lymphoblastic leukemia (29.0%) and acute myeloid leukemia (4.0%). Institutional vacci- nation protocols for cancer survivors, hematopoietic stem cells, and solid organ recipient chil- dren were developed and presented. There is a need to prepare national guidelines for vaccinating children with altered immunocompetence. Sharing vaccination practices by multidisciplinary vaccination units might increase and provide knowledge to develop national policies.


Effect of acupuncture on the TLR2/4-NF-κB signalling pathway in a rat model of traumatic brain injury.
To study the effect of acupuncture on the TLR2/4-NF-κB signalling pathway in the cortex of Sprague-Dawley rats following traumatic brain injury (TBI), and investigate the possible mechanism underlying the effects of acupuncture on scar repair. TBI was established using Feeney's free-falling epidural percussion model. In total, 108 rats were randomly divided into a normal group (n=18), untreated TBI model group (TBI group, n=36) and manual acupuncture-treated TBI group (TBI+MA, n=36). Each group of rats was subdivided into three time groups: 3-day (3d), 7-day (7d) and 14-day (14d). No treatment was given to rats in the normal and TBI groups. The TBI+MA group received manual acupuncture at GV20, GV26, GV16 through GV15, and bilateral LI4. mRNA expression of TLR2, TLR4, NF-κB and protein in the rat cortices was quantified using real-time fluorescence quantitative polymerase chain reaction (qPCR) and Western blot analyses. The modified neurological severity score (mNSS) scores of the TBI+MA group were improved compared with baseline scores 12 hours after modelling, and improved at 7d and 14d compared with the TBI group (P<0.05), while the score of the TBI group did not improve until 14d compared to baseline. mRNA and protein expression of TLR2, TLR4 and NF-κB in the TBI group were higher than the normal group at 3d (P<0.05), reached a peak at 7d, then began to decrease at 14d. mRNA and protein expression of TLR2, TLR4 and NF-κB were higher in the TBI+MA group compared with the TBI group at 3d (P<0.05), were significantly down-regulated at 7d (P<0.01), and decreased to normal levels at 14d. Acupuncture has a bidirectional regulatory effect on the TLR2/4-NF-κB signalling pathway-related genes TLR2, TLR4 and NF-κB in the TBI rat cortex, promoting their expression in the early stage and inhibiting it in the later stage.


Analysis of oilseed rape stem weevil chemical control using a damage rating scale.
Rape stem weevil (Ceutorhynchus napi Gyll.) and cabbage stem weevil (C. pallidactylus Marsh.) can cause significant yield losses to oilseed rape (Brassica napus L.), and chemical control is often needed to protect crops from these pests. The efficacy of six insecticides, chlorpyriphos + cypermethrin, bifenthrin, alpha-cypermethrin, pirimiphos-methyl, thiacloprid and tau-fluvalinate, was tested in a 4 year field trial. Besides the standard efficacy analysis expressed through the number of larvae per stem, a damage rating scale was introduced and modelled using a regression model for ordinal categorical data. Compared with the control, expressed through damage rating and larval number, treatments with chlorpyriphos + cypermethrin and bifenthrin, showed higher efficacy in the control of stem weevils compared with alpha-cypermethrin and pirimiphos-methyl. The lowest efficacy was observed in treatments with tau-fluvalinate and thiacloprid. This study showed that a combined efficacy evaluation expressed through both damage rating scale and the count of larvae, supported by an ordinal regression model for data analysis, is indispensable for obtaining accurate results. © 2017 Society of Chemical Industry.


Pancreatic exocrine secretion. Release of gastrin and cholecystokinin in response to bombesin in pigs.
The objective of this study was to determine whether bombesin can stimulate pancreatic exocrine secretion and release of cholecystokinin-33 and gastrin in pigs. Bombesin (1 microgram/kg-hr) was infused intravenously in six conscious piglets for one hour, and pancreatic secretions, plasma, and serum were collected. Cholecystokinin-33 and gastrin were measured by specific radioimmunoassays. Pancreatic secretory volume increased from a basal value of 1.5 +/- 0.5 mL/15 min to 10.4 +/- 1.7 mL/15 min. Pancreatic protein output increased from basal values of 7.9 +/- 2.4 mg/15 min to 88.9 +/- 25.4 mg/15 min. Plasma cholecystokinin-33 increased significantly from a basal concentration of 130 +/- 40 pg/mL to 275 +/- 100 pg/mL, as did gastrin (40.7 pg/mL to 200 +/- 27 pg/mL). Our findings demonstrated that bombesin stimulates exocrine pancreatic secretion in pigs; we conclude that this stimulation is due largely to the release of cholecystokinin.


Tubal sterilisation without consent: a case report.
Tubal sterilisation is practised all over the world. This safe, easy and highly effective, long-term method for birth control can be carried out during the hospital stay for either a normal delivery or caesarean section. In India, female sterilisation accounts for 37.3% of all methods of family planning.


America and the age of genocide: labeling a third-party conflict "genocide" decreases support for intervention among ingroup-glorifying Americans because they down-regulate guilt and perceived responsibility to intervene.
Drawing on research on the collapse of compassion and group processes and interrelations, four experiments investigated how labeling a conflict "genocide" affects distant bystanders' support for intervention. The genocide label (compared with no label or the label "not a genocide") weakened Americans' support for intervention in a crisis analogous to Darfur. Ingroup glorification moderated this effect such that the genocide label decreased support at high levels of glorification (Studies 1-3). Ingroup attachment, if anything, moderated such that the genocide label increased support at high levels of attachment (Studies 1 and 3). Importantly, the effects occurred even when controlling for conservatism (Studies 1 and 3), gender, religion, military affiliation, and level of education (Study 2). Decreases in anticipated guilt over possible nonintervention (Studies 1 and 3) among high glorifiers, and a subsequent decrease in perceived obligation to intervene (Study 3), mediated the effect of the genocide label on support for intervention.


Electrochemical Synthesis of Dihydrobenzofurans and Evaluation of Their Insect Antifeedant Activities.
Electrochemically synthesized dihydrobenzofurans were evaluated for their insect antifeedant activities against phytophagous insects. They were prepared through the coupling reactions of various alkenes with a phenoxy cation generated by oxidation near the cathode in the electrolytic reaction. The insect antifeedant activities of these synthetic dihydrobenzofurans were evaluated in the common cutworm (Spodoptera litura) and diamond back moth (Plutella xylostella) with the dual choice leaf disk bioassay method. The insect antifeedant activities of most of the acetophenone-type dihydrobenzofurans were strong, while those of derivatives with a t-butyl group were weaker. The biological activities in insect species differed with the structural features of the compounds.


Can Preoperative CT Scans Be Used to Predict Facial Nerve Stimulation Following CI?
1) To determine the ability of preoperative computed tomography (CT) to predict facial nerve stimulation (FNS) after cochlear implantation (CI). 2) To recognize the limitations of CT in predicting FNS. Patient control study. Tertiary care academic medical center. Adult patients with CI from 2003 to 2015. Patients with severe FNS (n = 4) were compared with randomly selected CI patients (n = 28). Three blinded reviewers evaluated preoperative temporal bone CT scans to measure the distance from the labyrinthine segment of the facial nerve to the basal turn of the cochlea and attempted to predict whether or not the subject had FNS after CI. In total, 32 CT scans were evaluated representing 49 ears that underwent CI.The distances (mm) measured from the labyrinthine segment of the facial nerve to the basal turn of the cochlea in both the axial (0.3 ± 0.3 versus 0.6 ± 0.3) and coronal (0.4 ± 0.2 versus 0.6 ± 0.2) orientation were significantly different between the two groups (p = 0.0001 and p = 0.0034) respectively. The intraclass correlation coefficient demonstrated good (K > 0.7) reviewer correlation in both the reviewers' measurements and predictions. The sensitivity, specificity, positive predictive value, and negative predictive value for preoperative CT scans to predict FNS were 38.5, 85.1, 19.2, and 93.8% respectively. The reviewers were 23% accurate in predicting FNS. Based on a blinded retrospective patient-control study, CT scan measurements show a significantly reduced distance between the labyrinthine facial nerve and the basal turn of the cochlea in patients with FNS. However, it is difficult to predict who will have FNS based on these measurements.


The optimal timing of hemoperfusion component in combined hemodialysis-hemoperfusion treatment for uremic toxins removal.
The combination of hemodialysis-hemoperfusion (HDHP) has been proved to be superior to hemodialysis (HD) in eliminating uremic toxins. There are two methods of combination of HD and HP: the HP regime is utilized during the first two-hour an early HP conducted HDHP (EHDHP) or the last two-hour late HP conducted HDHP (LHDHP) of 4 h regular HD session. The present study was to compare these two methods in uremic toxins removal. Twenty adult chronic HD patients were enrolled in this self-control method study. The patients were randomized to receive one session of EHDHP or LHDHP. Two weeks later, the dialysis modalities were switched. The reduction ratio (RR) of targeted uremic toxins for each session was assessed. Both EHDHP and LHDHP showed a significant removal of small water-soluble solutes, middle-sized toxins and cytokines as well (p < 0.05). There were no significant differences between two methods in RR of small water-soluble solutes, like urea and creatinine. For middle-sized molecules and cytokines, such as PTH, β(2)-M, IL-1, IL-6, and TNF-α, the RR was markedly increased in LHDHP than that in EHDHP (p < 0.05). LHDHP showed no more intradialytic events than EHDHP. The combination of HD and HP in the last two hours in one hemodialysis session had more effect on eliminating middle-sized toxins and cytokines.


Inhibition of deoxyribonucleic acid gyrase: effects on nucleic acid synthesis and cell division in Escherichia coli K-12.
Mutants of Escherichia coli resistant to the antibiotic clorobiocin are also coumermycin resistant, and the mutation to resistance in at least one mutant was mapped near gyrB. We conclude, therefore, that clorobiocin inhibits deoxyribonucleic acid gyrase, and the drug was used to probe the role of this enzyme in vivo. Deozyribonucleic acid synthesis was preferentially inhibited but not completely blocked by the antibiotic. Transcription and cell division were also markedly affected. However, unlike other inhibitors of deoxyribonucleic acid synthesis, clorobiocin failed to induce the synthesis of protein X, the recA gene product. In mutants resistant to clorobiocin the replication velocity was unaffected, but initiation of deoxyribonucleic acid synthesis appeared to be delayed. We conclude that deoxyribonucleic acid gyrase, and hence the supercoiled structure of the chromosome, is important for transcription, normal initiation of deoxyribonucleic acid replication, and cell division. The possible role of deoxyribonucleic acid gyrase in the elongation of replication forks is also discussed.


Ultrastructural studies of the ovary of Palaeococcus fuscipennis (Burmeister) (Insecta, Hemiptera, Coccinea: Monophlebidae).
Ovaries of Palaeocoocus fuscipennis are composed of about 100 telotrophic ovarioles that are devoid of terminal filaments. In the ovariole a tropharium ( = trophic chamber) and vitellarium can be distinguished. The tropharium contains 7 trophocytes. A single oocyte develops in the vitellarium. The oocyte is surrounded by follicular cells that do not undergo diversification into subpopulations. The obtained results are discussed in a phylogenetic context.


[Experimental study on culture of human mesenchymal stem cells from cord blood using autologous serum].
In this study, we collected human umbilical cord blood and separated the autologous serum and clear cells. Then the human mesenchymal stem cells were cultured with the use of autologous serum and fetal serum, respectively. By identifing surface markers with flow cytometry and testing the osteogenic, adipogenic and cardiomyocytes differentiation capacity of the cells, we affirmed that the cultured cells are mesenchymal stem cells. Among 21 cases of umbilical cord blood, 11 cases using fetal bovine serum and 15 cases using autologous successfully cultured mesenchymal stem cells from umbilical cord blood. The UCB-MSCs did not express CD34 and CD45, but they expressed CD29, CD44 and CD105, and they were successfully induced into osteogenic cells, adipogenic cells and cardiomyocytic-like cells. All of these suggest that using autologous serum to culture UCB-MSCs is better than using fetal bovine serum, and the autologous serum cultured UCB-MSCs can also be induced into osteogenic cells, adipogenic cells cardiomycyote-like cells.


Effect of Allium sativum and Allium cepa oils on different stages of Boophilus annulatus.
The study is aimed to investigate the acaricidal effect of Allium sativum (garlic) and Allium cepa (onion) oils on different stages of Boophilus annulatus hard tick. Engorged B. annulatus females were collected from naturally infected cattle. A number of engorged ticks were incubated at 28 °C and 85 % relative humidity to lay eggs, which were incubated to obtain larvae that were used in the study. The used garlic and onion oils were prepared by steam distillation and were analyzed by gas chromatography. These oils were dissolved in ethanol, methanol alcohols, and, partially, in water. The oils were tested in different concentrations; 1, 2, 5, 10, and 20 %. These concentrations were applied on adult ticks by adult immersion test; on larvae by larval immersion technique and on eggs. The 20, 10, and 5 % of garlic oil dilutions in ethanol and methanol killed all adult ticks and larvae within 24 h. Similar results were obtained for 10 and 20 % garlic oil dissolved in water. The effect of 10 % aqueous solution of garlic oil on embryonated eggs was clear as its addition to these eggs led to their in ability to hatch, deformity in shape, and change in color. The 10 and 20 % onion oil in ethanol and methanol alcohols killed 76-86 % of the adult ticks within 72 h post-application. While, all larvae died within 24 h postsubjected to these two concentrations. These concentrations (10 and 20 %) of onion oil in water killed 56-80 % of the treated ticks. Moreover, 10 % aqueous solution of onion oil prevented hatching of embyonated eggs. We concluded that garlic and onion oils have acaricidal effect on all stages of B. annulatus at concentrations higher than 5 %. Only garlic oil could kill 100 % of adult ticks at concentrations from 5 % in alcohols.


Liquid Biopsy in NSCLC: An Investigation with Multiple Clinical Implications.
Tissue biopsy is essential for NSCLC diagnosis and treatment management. Over the past decades, liquid biopsy has proven to be a powerful tool in clinical oncology, isolating tumor-derived entities from the blood. Liquid biopsy permits several advantages over tissue biopsy: it is non-invasive, and it should provide a better view of tumor heterogeneity, gene alterations, and clonal evolution. Consequentially, liquid biopsy has gained attention as a cancer biomarker tool, with growing clinical applications in NSCLC. In the era of precision medicine based on molecular typing, non-invasive genotyping methods became increasingly important due to the great number of oncogene drivers and the small tissue specimen often available. In our work, we comprehensively reviewed established and emerging applications of liquid biopsy in NSCLC. We made an excursus on laboratory analysis methods and the applications of liquid biopsy either in early or metastatic NSCLC disease settings. We deeply reviewed current data and future perspectives regarding screening, minimal residual disease, micrometastasis detection, and their implication in adjuvant and neoadjuvant therapy management. Moreover, we reviewed liquid biopsy diagnostic utility in the absence of tissue biopsy and its role in monitoring treatment response and emerging resistance in metastatic NSCLC treated with target therapy and immuno-therapy.


Optimization strategies for production of mammalian embryos by nuclear transfer.
In order to optimize each of the individual steps in the nuclear transfer procedure, we report alternative protocols useful for producing recipient cytoplasts and for improving the success rate of nuclear transfer embryos in cattle, rhesus monkey, and hamster. Vital labeling of maternal chromatin/spindle is accomplished by long wavelength fluorochromes Sybr14 and rhodamine labeled tubulin allowing constant monitoring and verification during enucleation. The use of Chinese hamster ovary (CHO) donor cells expressing the viral influenza hemagglutinin fusion protein (HA-300a+), to adhere and induce fusion between the donor cells and enucleated cow, rhesus and hamster oocytes was examined. Cell surface hemagglutinin was activated with trypsin prior to nuclear transfer and fusion was induced by a short incubation of a newly created nuclear transfer couplet at pH 5.2 at room temperature. Donor cell cytoplasm was dynamically labeled with CMFDA, or further transfected with the green fluorescence protein (GFP) gene, so that fusion could be directly monitored using live imaging. High rates of fusion were observed between CHO donor cells and hamster (100%), rhesus (100%), and cow recipient cytoplasts (81.6%). Live imaging during fusion revealed rapid intermixing of cytoplasmic components between a recipient and a donor cell. Prelabeled donor cytoplasmic components were uniformly distributed throughout the recipient cytoplast, within minutes of fusion, while the newly introduced nucleus remained at the periphery. The fusion process did not induce activation as evidenced by unchanged distribution and density of cortical granules in the recipient cytoplasts. After artificial activation, the nuclear transfer embryos created in this manner were capable of completing several embryonic cell divisions. These procedures hold promise for enhancing the efficiency of nuclear transfer in mammals of importance for biomedical research, agriculture, biotechnology, and preserving unique, rare, and endangered species.


[A study of factors associated to non-pharmacological treatment delivered by the Family Health Program in Ventosa, Belo Horizonte, to carriers of diabetes mellitus].
Diabetes Mellitus is turning into one of the main public health problems in Brazil. Many factors may contribute to the low adherence to treatment. The objective of this study was to investigate the association between educational and social-economic factors and factors related to the health, the perception of disease, social support and adherence to non-pharmacological treatment of carriers of diabetes mellitus assisted in the family health program of the Health Center in Ventosa, Belo Horizonte. Data were collected in interviews with 164 diabetics who were waiting for assistance in the health center. Univariate analysis showed significant association between adhesion to the non-pharmacological treatment and low income; place of residence; knowledge about consequences; motivation with regard to the treatment; making part of a diabetics group and orientation received from the nurse and the physiotherapist. Multivariate analysis showed the variables " treatment motivation" and "part of some diabetics group", to be statistically significant as refers to the total adhesion (p <0.05). The variables "knowledge about consequences " and " place of residence--high risk area" were statistically significant as refers to the total adhesion (p < 0.06), however presenting an inverse relation.


Multifocal corneal ablation for hyperopic presbyopes.
To investigate the clinical results of multifocal wavefront-guided corneal ablation with a central near zone in hyperopic presbyopes. Twenty-seven patients (54 eyes) were analyzed retrospectively. Monocular near and distance uncorrected visual acuity (UCVA) and binocular near and distance UCVA were measured preoperatively, and the near addition power and the manifest refraction were obtained. Measurements of spherical aberration, coma, total higher order aberration, and contrast sensitivity for high and low photopic conditions were performed. All patients received multifocal corneal ablations with a central near zone using the VISX CustomVue System and were examined at 1 month (48 eyes), 2 months (42 eyes), and 6 months (28 eyes) after surgery. Of 14 patients, 9 (64.3%) achieved both 0.8 or better in binocular distance UCVA and 0.65 or better in binocular near UCVA, and the spherical equivalent refraction and near addition power decreased from +1.16+/-0.82 diopters (D) to +0.33+/-0.45 D and from 2.35+/-0.44 D to 1.42+/-0.62 D at 6 months after surgery, respectively. No significant changes were noted in total higher order aberration and contrast sensitivity, whereas spherical aberration coefficients (Z-12) changed from positive to negative. Multifocal wavefront-guided refractive surgery in presbyopia offered effective correction in hypermetropia and satisfactory near vision without significant increase in comeal total higher order aberration or significant decrease in contrast sensitivity.


Novel Ceramic-Grafted Separator with Highly Thermal Stability for Safe Lithium-Ion Batteries.
The separator is a critical component of lithium-ion batteries (LIBs), which not only allows ionic transport while it prevents electrical contact between electrodes but also plays a key role for thermal safety performance of LIBs. However, commercial separators for LIBs are typically microporous polyolefin membranes that pose challenges for battery safety, due to shrinking and melting at elevated temperature. Here, we demonstrate a strategy to improve the thermal stability and electrolyte affinity of polyethylene (PE) separators. By simply grafting the vinylsilane coupling reagent on the surface of the PE separator by electron beam irradiation method and subsequent hydrolysis reaction into the Al3+ solution, an ultrathin Al2O3 layer is grafted on the surface of the porous polymer microframework without sacrificing the porous structure and increasing the thickness. The as-synthesized Al2O3 ceramic-grafted separator (Al2O3-CGS) shows almost no shrinkage at 150 °C and decreases the contact angle of the conventional electrolyte compared with the bare PE separator. Notably, the full cells with the Al2O3-CGSs exhibit better cycling performance and rate capability and also provide stable open circuit voltage even at 170 °C, indicating its promising application in LIBs with high safety and energy density.


How thermal stress alters the confinement of polymers vitrificated in nanopores.
Understanding and controlling the glass transition temperature (Tg) and dynamics of polymers in confined geometries are of significance in both academia and industry. Here, we investigate how the thermal stress induced by a mismatch in the coefficient of thermal expansion affects the Tg behavior of polystyrene (PS) nanorods located inside cylindrical alumina nanopores. The size effects and molecular weight dependence of the Tg are also studied. A multi-step relaxation process was employed to study the relationship between thermal stress and cooling rate. At fast cooling rates, the imparted thermal stress would overcome the yield stress of PS and peel chains off the pore walls, while at slow cooling rates, chains are kept in contact with the pore walls due to timely dissipation of the produced thermal stress during vitrification. In smaller nanopores, more PS chains closely contact with pore walls, then stronger internal thermal stress would be generated between core and shell of PS nanorod, which results in a larger deviation between two Tgs. The core part of PS shows lower Tg than bulk value, which can induce faster dynamics in the center region. A complex and important role stress plays is supposed in complex confinement condition, e.g., in nanopores, during vitrification.


Global surgery: current evidence for improving surgical care.
The field of global surgery is undergoing rapid transformation, owing to several recent prominent reports positioning it as a cost-effective means of relieving global disease burden. The purpose of this article is to review the recent advances in the field of global surgery. Efforts to grow the global surgical workforce and procedural capacity have focused on innovative methods to increase surgeon training, enhance international collaboration, leverage technology, optimize existing health systems, and safely implement task-sharing. Computer modeling offers a novel means of informing policy to optimize timely access to care, equitably promote health and financial protection, and efficiently grow infrastructure. Tools and checklists have recently been developed to enhance data collection and ensure methodologically rigorous publications to inform planning, benchmark surgical systems, promote accurate modeling, track key health indicators, and promote safety. Creation of institutional partnerships and trainee exchanges can enrich training, stimulate commitment to humanitarian work, and promote the equal exchange of ideas and expertise. The recent body of work creates a strong foundation upon which work toward the goal of universal access to safe, affordable surgical care can be built; however, further collection and analysis of country-specific data is necessary for accurate modeling and outcomes research into the efficacy of policies such as task-sharing is greatly needed.


Adenosquamous carcinoma of the bile duct: a population-based study.
Adenosquamous carcinoma (ASC) of the bile duct is a rare diagnosis with poorly understood clinicopathological characteristics and disease progression, so identification of the features associated with ASC patient survival is warranted. A population cohort study was performed using prospectively extracted data from the Surveillance, Epidemiology and End Results (SEER) database for patients with histological diagnoses of ASC of the bile duct from 1973 to 2013. A total of 106 patients with ASC of the bile duct were included (mean age 68.1±13.5 years). Lesions from 58 patients were in the extrahepatic bile duct and 34 were located at the ampulla of Vater. Fifty-seven patients were categorized with a regional stage, 15 had localized disease, and 30 had distant disease. Most (60.4%) patients received cancer-directed surgery, and radiation was performed in 14.1% of cases. The 1-year, 2-year, and 5-year overall survival (OS) for patients with ASC of the bile duct was 30.1%, 11.3%, and 3.7%, respectively. Cancer-directed surgery offered 10 additional months of OS for patients with ASC of the bile duct and median OS was 14.0, 6.0, and 6.0 months for ampulla of Vater, extrahepatic bile duct, and intrahepatic bile duct cases, respectively. A multivariate Cox analysis showed that lesions in the ampulla of Vater (HR=0.51, 95% CI 0.26-0.99) and having surgery (HR=0.34, 95% CI 0.14-0.81) were independent protective prognostic factors for these patients. Cancer-directed surgery and a primary lesion site of the ampulla of Vater may suggest favorable prognosis for patients with ASC of the bile duct.


Does training respiratory physicians in clinical respiratory physiology and interpretation of pulmonary function tests improve core knowledge?
Lung function tests have a major role in respiratory medicine. Training in lung function tests is variable within the European Union. In this study, we have shown that an internship in a lung function tests laboratory significantly improved the technical and diagnostic skills of French respiratory trainees.


Prion protein remodelling confers an immediate phenotypic switch.
In a variety of systems, proteins have been linked to processes historically limited to nucleic acids, such as infectivity and inheritance. These atypical proteins, termed prions, lack sequence homology but are collectively defined by their capacity to adopt multiple physical and therefore functional states in vivo. Newly synthesized prion protein generally adopts the form already present in the cell, and this in vivo folding bias directs the near faithful transmission of the corresponding phenotypic state. Switches between the prion and non-prion phenotypes can occur in vivo; however, the fate of existing protein during these transitions and its effects on the emergence of new traits remain major unanswered questions. Here, we determine the changes in protein-state that induce phenotypic switching for the yeast prion Sup35/[PSI(+)]. We show that the prion form does not need to be specified by an alternate misfolding pathway initiated during Sup35 synthesis but instead can be accessed by mature protein. This remodelling of protein from one stable form to another is accompanied by the loss of Sup35 activity, evoking a rapid change in cellular phenotype within a single cell cycle.


Modeling the fate of antibiotic resistance genes and class 1 integrons during thermophilic anaerobic digestion of municipal wastewater solids.
This study investigated the use of thermophilic anaerobic digestion for removing antibiotic resistance genes (ARGs) from residual municipal wastewater solids. Four laboratory-scale anaerobic digesters were operated in 8-day batch cycles at temperatures of 40, 56, 60, and 63 °C. Two tetracycline resistance genes (tet(W) and tet(X)), a fluoroquinolone resistance gene (qnrA), the integrase gene of class 1 integrons (intI1), 16S rRNA genes of all Bacteria, and 16S rRNA genes of methanogens were quantified using real-time quantitative PCR. ARG and intI1 quantities decreased at all temperatures and were described well by a modified form of the Collins-Selleck disinfection kinetic model. The magnitudes of Collins-Selleck kinetic parameters were significantly greater at thermophilic temperatures compared to 40 °C, but few statistically significant differences were observed among these parameters for the thermophilic anaerobic digesters. This model allows for the direct comparison of different operating conditions (e.g., temperature) on anaerobic digestion performance in mitigating the quantity of ARGs in wastewater solids and could be used to design full-scale anaerobic digesters to specifically treat for ARGs as a "pollutant" of concern.


Electrophysiological correlates of flicker-induced color hallucinations.
In a recent study, Becker and Elliott [Becker, C., & Elliott, M. A. (2006). Flicker induced color and form: Interdependencies and relation to stimulation frequency and phase. Consciousness & Cognition, 15(1), 175-196] described the appearance of subjective experiences of color and form induced by stimulation with intermittent light. While there have been electroencephalographic studies of similar hallucinatory forms, brain activity accompanying the appearance of hallucinatory colors was never measured. Using a priming procedure where observers were required to indicate the presence of one of eight target colors we compared electrophysiological correlates of hallucinatory color with brain states associated with other visual phenomena. Different target colors were accompanied by different patterns of EEG activation. However, in general, we found that the appearance of hallucinatory colors is preceded by a power decrease in the lower alpha band alongside an increase in gamma band frequencies. We argue that decreasing activity in the lower alpha band acts as a gating mechanism, inducing a switch in perception between different colors. The increasing gamma activation may correlate with the formation of a coherent conscious percept.


Promoting Photosynthetic Production of Dammarenediol-II in Chlamydomonas reinhardtii via Gene Loading and Culture Optimization.
Ginsenosides are major bioactive compounds found in Panax ginseng that exhibit various pharmaceutical properties. Dammarenediol-II, the nucleus of dammarane-type ginsenosides, is a promising candidate for pharmacologically active triterpenes. Dammarenediol-II synthase (DDS) cyclizes 2,3-oxidosqualene to produce dammarenediol-II. Based on the native terpenoids synthetic pathway, a dammarane-type ginsenosides synthetic pathway was established in Chlamydomonas reinhardtii by introducing P. ginseng PgDDS, CYP450 enzyme (PgCYP716A47), or/and Arabidopsis thaliana NADPH-cytochrome P450 reductase gene (AtCPR), which is responsible for producing dammarane-type ginsenosides. To enhance productivity, strategies such as "gene loading" and "culture optimizing" were employed. Multiple copies of transgene expression cassettes were introduced into the genome to increase the expression of the key rate-limiting enzyme gene, PgDDS, significantly improving the titer of dammarenediol-II to approximately 0.2 mg/L. Following the culture optimization in an opt2 medium supplemented with 1.5 mM methyl jasmonate under a light:dark regimen, the titer of dammarenediol-II increased more than 13-fold to approximately 2.6 mg/L. The C. reinhardtii strains engineered in this study constitute a good platform for the further production of ginsenosides in microalgae.


Granulocytes cause reperfusion ventricular dysfunction after 15-minute ischemia in the dog.
Regional ventricular dysfunction (the stunned myocardium) persists for several hours after 15 minutes of ischemia and reperfusion in the dog. Superoxide-radical-induced damage appears to be one of the mechanisms of this injury. We tested whether granulocytes were a direct source of injury in the stunned myocardium in the 15-minute ischemia dog model. Regional function during agranulocytic extracorporeal coronary perfusion (using Leukopak filters) with ischemia and reperfusion was compared with function during a second period of ischemia and reperfusion after removal of the filters (granulocytopenia). Flow reduction and reperfusion flow, preload, afterload, and inotropic stimulation were the same during agranulocytic and granulocytopenic perfusion. During agranulocytic perfusion, stunning did not occur (greater than 100% of preischemic function during reperfusion), but when the filters were removed and about 10% of the normal granulocyte count was present, stunning occurred with only 76% return of function at 60 minutes of reperfusion (p less than 0.01). A second series of studied animals with extracorporeal perfusion and granulocyte replete perfusion all had less than 75% return of regional function, indicating that the agranulocytic perfusion and not the extracorporeal aspects of the experiment prevented stunning. We conclude that granulocytes are the direct source of the injury in stunned myocardium and apparently the main source of superoxide in the 15-minute ischemia dog model. Other possible granulocyte-related mechanisms of reperfusion injury include capillary no-reflow, causing microvascular ischemia and degranulation leading to enzyme-induced damage.


Predictors of Postoperative Atrial Fibrillation after Isolated On-Pump Coronary Artery Bypass Grafting in Patients ≥60 Years Old.
Postoperative atrial fibrillation (POAF) after coronary artery bypass grafting (CABG) is one of the most common complications that can contribute to an increased risk of mortality, particularly in older patients. The identification of predictors of POAF after CABG could impact patient care. This study aims to determine the risk factors of POAF in patients ≥60 years old who underwent isolated on-pump coronary artery bypass grafting (ONCAB) in order to provide a basis for the prevention and treatment of POAF after ONCAB. Between October 2011 and November 2015, a total of 304 patients ≥60 years old underwent isolated ONCAB in our department. The patients were divided into 2 groups, the AF group and the non-AF group, according to the occurrence of POAF. A retrospective analysis was performed on the general characteristics and perioperative data of the patients. Logistic regression analysis was used to identify the predictors of POAF after ONCAB in patients ≥60 years old. The incidence of POAF after ONCAB in patients ≥60 years old was 23.36% (71/304). Statistically significant differences were observed in patients' age, history of hypertension, left atrium diameter, European System for Cardiac Operative Risk Evaluation (EuroSCORE) I and II, and the highest level of serum creatinine after operation (all P < .05). Logistic regression analyses showed that a history of hypertension (OR = 2.575, CI 1.208-5.488, P = .014), left atrium diameter (OR = 1.105, CI 1.047-1.167, P = .000) and EuroSCORE I score (OR = 1.132, CI 1.001-1.279, P = .048) were independent risk factors for POAF after ONCAB in patients ≥60 years old. The occurrence of POAF after isolated ONCAB in patients ≥60 years old was affected by many risk factors; a history of hypertension, the left atrium diameter and the EuroSCORE I score were all predictors of POAF.


Intrinsic Property Forecast Index (iPFI) as a Rule of Thumb for Medicinal Chemists to Remove a Phototoxicity Liability.
Phototoxicity occurs when UV irradiation causes otherwise benign compounds to become irritant, sensitizers, or even genotoxic. This toxicity is particularly a concern after topical application and in dermatological programs where skin irritation can be incompatible with the desired therapeutic outcome. This brief article establishes that the intrinsic property forecast index (iPFI) can be used to evaluate the probability of a compound being phototoxic and gives medicinal chemists a practical tool to handle this liability.


[The role of morphometry in the cytology of pleomorphic adenomas of the salivary glands].
A morphometric analysis was performed on cytologic smears (FNAB) of 5 cases of pleomorphic adenoma that recurred after the first diagnosis and on 5 ordinary pleomorphic adenoma. The considered parameters were as follows: nuclear area, perimeter, maximum diameter, feret x, feret y, F shaped and D circle. A statistical analysis was performed between the two groups for all the considered parameters using a two-sided Student's t-test for independent samples. The test showed significant differences of the parameter nuclear area. The authors conclude that a morphometric assessment may bring a significant contribution in the knowledge and prediction of the outcome of pleomorphic adenoma.


Static and dynamic pupillary features in graves' ophthalmopathy.
Pupillary characteristics may be affected in patients with Graves' ophthalmopathy (GO). However, small changes cannot be observed with clinical examination. Ophthalmologists and optometrists should be aware that dynamic pupillometry may have an important place in the quantitative assessment of pupillary characteristics in this patient population. The aim of this study was to compare the static and dynamic pupillary responses of hyperthyroid and euthyroid GO patients and healthy control subjects. The study enrolled 20 hyperthyroid patients with GO (Group 1), 20 euthyroid patients with GO (Group 2) and 40 control subjects with normal thyroid function tests and no known illness (Group 3). Following detailed ophthalmological examination, static and dynamic pupillometry measurements were performed. Dynamic pupillometry measurements including resting diameter, amplitude of pupil contraction, latency of pupil contraction, duration of pupil contraction, latency of pupil dilation, duration of pupil dilation, and velocity of pupil dilation were undertaken. Static pupillometry measurements including scotopic, mesopic, low-photopic, and high-photopic pupil diameters (PD) were undertaken. Data from the right eyes of the participants were used for statistical analysis. Regarding the dynamic pupillary measurements, the latency of pupil contraction value was significantly higher (p = 0.007), and the velocity of pupil dilatation was significantly lower (p = 0.004) in Groups 1 and 2 compared to the Group 3. In static pupillary measurements, there were statistically significant differences between the GO group (Group 1 and 2) and Group 3 with regard to scotopic (p = 0.002), mesopic (p = 0.002), and low-photopic PD (p = 0.001). Scotopic, mesopic and low photopic PD, latency of pupil contraction and velocity of pupil dilatation values were significantly different in both hyperthyroid and euthyroid GO patients when compared to healthy control subjects.


Transient hypoxemia induced by cortical electrical stimulation: A mapping study in 75 patients.
To identify which cortical regions are associated with direct electrical stimulation (DES)-induced alteration of breathing significant enough to impair pulse oximetry (SpO2). Evolution of SpO2 after 1,352 DES was analyzed in 75 patients with refractory focal epilepsy who underwent stereo-EEG recordings. For each DES, we assessed the change in SpO2 from 30 seconds prior to DES onset to 120 seconds following the end of the DES. The primary outcome was occurrence of stimulation-induced transient hypoxemia as defined by decrease of SpO2 ≥5% within 60 seconds after stimulation onset as compared to pre-DES SpO2 or SpO2 nadir <90% during at least 5 seconds. Localization of the stimulated contacts was defined according to MarsAtlas brain parcellation and Freesurfer segmentation. A stimulation-induced transient hypoxemia was observed after 16 DES (1.2%) in 10 patients (13%), including 6 in whom SpO2 nadir was <90%. Among these 16 DES, 7 (44%) were localized within the perisylvian cortex. After correction for individual effects and the varying number of DES contributed by each person, significant decrease of SpO2 was significantly associated with the localization of DES (p = 0.019). Though rare, a significant decrease of SpO2 could be elicited by cortical direct electrical stimulation outside the temporo-limbic structures, most commonly after stimulation of the perisylvian cortex.


Epigenetic modulation of PGC-1α activity by GCN5 inhibitors: WO2010007085.
The transcriptional peroxisome proliferator-activated receptor γ (PPARγ) co-activator PGC-1α plays a central role in the regulation of cellular energy metabolism. Among the wide range of its activities, PGC-1α controls mitochondrial biogenesis and function and is one of the main factors involved in hormonal and nutrient regulation of hepatic gluconeogenesis. PGC-1α is present in a multiprotein complex, and its activity can also be modulated through epigenetic modifications. In particular, it is directly acetylated by the HAT enzyme general control nonderepressible 5 (GCN5), resulting in a transcriptionally inactive protein that relocalizes from promoter regions to nuclear foci, whereas it is deacetylated by SIRT1 at multiple lysine sites, with a subsequent increase in its activity leading to induction of liver gluconeogenic gene transcription. Thus, both GCN5 and SIRT1 may be pharmacological targets to regulate the activity of PGC-1α, providing a potential treatment for metabolic disorders in which hepatic glucose output is altered.


Relapsing neuromyelitis optica in an adolescent girl.
Early differentiation of neuromyelitis optica spectrum disorder (NMO-SD) from multiple sclerosis (MS) is of paramount importance as NMO-SD (especially relapsing variant) has more severe morbidity than MS. We describe a case of an adolescent girl who presented with repeated episodes of optic neuritis over a period of 4 years with normal brain MRI scans. She was treated initially as relapsing remitting MS, before showing clinical evidence of transverse myelitis (TM), and eventually being diagnosed as NMO-SD. Pulse intravenous methyl prednisolone along with immunosuppressive therapy led to remission of her disease. However, delay in diagnosis as NMO-SD led to visual disability in the left eye. Therefore, in young patients with recurrent optic neuritis and normal brain MRI, it may be prudent to get spinal MRI done to look for TM, even when asymptomatic. In addition, we should keep a low threshold for requesting aquaporin-4 antibody testing in these patients.


Influence of tumor vaccines on graft versus tumor activity and graft versus host disease in allogeneic bone marrow transplantation.
Powerful immunologically-mediated antitumor efforts can be observed in allogeneic hematopoietic stem cell transplantation. In the absence of specific immune interventions, this graft versus tumor effect is closely associated with graft versus host disease. In the work summarized here, the influence of cellular tumor vaccines on graft versus tumor activity and graft versus host disease is examined in a murine model of MHC-matched, minor histocompatibility antigen-mismatched bone marrow transplantation. The experiments have generated the following conclusions. First, complex cellular vaccines, which include recipient minor histocompatibility antigens, when administered to allogeneic donors generate powerful graft versus tumor effects but also induce unacceptable exacerbations of graft versus host disease. Second, cellular tumor vaccines, which contain recipient minor histocompatibility antigens, can be administered to transplant recipients after transplant without significant exacerbation of GVHD and with retention of clinically significant graft versus tumor effects. Third, immunization of donors with molecularly defined tumor-associated antigens, which are not recipient minor histocompatibility antigens, can be coupled with post-transplant immunization of recipients with cellular vaccines without exacerbation of GVHD.


Cross-Platform DNA Encoding for Single-Cell Imaging of Gene Expression.
Integration of imaging data across different molecular target types can provide in-depth insight into cell physiology and pathology, but remains challenging owing to poor compatibility between target-type-specific labeling methods. We show that cross-platform imaging analysis can be readily achieved through DNA encoding of molecular targets, which translates the molecular identity of various target types into a uniform in situ array of ssDNA tags for subsequent labeling with complementary imaging probes. The concept was demonstrated through multiplexed imaging of mRNAs and their corresponding proteins with multicolor quantum dots. The results reveal heterogeneity of cell transfection with siRNA and outline disparity in RNA interference (RNAi) kinetics at the level of both the mRNA and the encoded protein.


Challenges in management of ST elevation myocardial infarction during COVID-19 pandemic.
The coronavirus disease-2019 (COVID-19) pandemic has brought unprecedented changes to our world and health-care system. Its high virulence and infectiousness directly infect people's respiratory system and indirectly disrupt our health-care infrastructure. In particular, ST elevation myocardial infarction (STEMI) is a clinical emergency emphasizes on the establishment of care system to minimize delay to reperfusion. As such, the impact of COVID-19 on STEMI care, ranging from disease severity, patient delay, diagnostic difficulty, triage to selection of reperfusion strategy and postoperative care, is immense. Importantly, not only we have to save our patients, but we must also need to protect all health-care workers and prevent environmental contamination. Otherwise, in-hospital transmission can quickly evolve into nosocomial outbreak with staff infection and quarantine which lead to health-care system collapse. In this article, we will discuss the challenges in various aspects of STEMI management during COVID-19, as well as the mitigation measures we can take to optimize outcome and our future.


Molecular Identification, Pathogenesis, and Life Cycle of Sarcocystis cruzi from Cattle (Bos taurus) in New Valley Governorate, Egypt.
Sarcocystis cruzi was identified by molecular methods from an intermediate host, cattle (Bos taurus), in El-Kharga, New Valley Governorate, Egypt, and its life cycle and pathogenicity were studied in the final host, dogs (Canis familiaris). 600 slaughtered cattle aged 6-8 years (480/120 males/females) were included. In addition, three laboratory-bred, coccidian-free puppies aged 2-3 months were fed infected bovine muscles to locate the definitive host and analyze sporogony. 18S rRNA-specific gene primers were used for DNA amplification from esophageal muscles. These polymerase chain reaction (PCR) amplicons were subjected to restriction fragment length polymorphism (RFLP) and molecular sequence analysis. Infection was detected in 78.8% (473/600; 95% CI, 75.56-82.11%). Histopathological examination of esophageal muscles showed oval- to spherical-shaped cysts, 96.7 μm wide by 326.9 μm long; cysts in cardiac muscles were ovoid and smaller. Infected puppies began shedding sporocysts in feces 7 days post-inoculation and showed distorted organ architecture, severe cellular damage, and inflammatory lesions in liver, kidney, esophagus, and stomach. Three oocysts with different shapes and sizes were identified. Partial 18S rRNA gene sequences of isolated New Valley sarcocysts were identical to S. cruzi isolated from different areas, verifying their genetic relatedness. Our analysis suggests that S. cruzi is the most prevalent in slaughtered cattle in New Valley Governorate, Egypt.


Protein Hydrolysate from Underutilized Legumes: Unleashing the Potential for Future Functional Foods.
Proteins play a vital role in human development, growth, and overall health. Traditionally, animal-derived proteins were considered the primary source of dietary protein. However, in recent years, there has been a remarkable shift in dietary consumption patterns, with a growing preference for plant-based protein sources. This shift has resulted in a significant increase in the production of plant proteins in the food sector. Consequently, there has been a surge in research exploring various plant sources, particularly wild, and underutilized legumes such as Canavalia, Psophocarpus, Cajanus, Lablab, Phaseolus, and Vigna, due to their exceptional nutraceutical value. This review presents the latest insights into innovative approaches used to extract proteins from underutilized legumes. Furthermore, it highlights the purification of protein hydrolysate using Fast Protein Liquid Chromatography. This review also covers the characterization of purified peptides, including their molecular weight, amino acid composition, and the creation of three-dimensional models based on amino acid sequences. The potential of underutilized legume protein hydrolysates as functional ingredients in the food industry is a key focus of this review. By incorporating these protein sources into food production, we can foster sustainable and healthy practices while minimizing environmental impact. The investigation of underutilized legumes offers exciting possibilities for future research and development in this area, further enhancing the utilization of plant-based protein sources.


Acquired toxoplasmosis accompanied by facial nerve palsy in an immunocompetent 5-year-old child.
Acquired toxoplasmosis, although relatively common in children, is usually asymptomatic but can also be clinically manifested by a benign and self-limited infectious mononucleosis-like syndrome. Neurological complications are very rare in immunocompetent children. The authors report a 5-year-old boy who presented with cervical lymphadenopathy because of acquired toxoplasmosis accompanied with unilateral facial nerve paralysis. Toxoplasma gondii DNA detection in blood by polymerase chain reaction, as well as elevated specific immunoglobulin M antibodies against it, established the diagnosis. Characteristic brain lesions on magnetic resonance imaging were absent and ophthalmologic examination revealed no inflammatory lesions in the retina and choroid. Treatment with pyrimethamine, sulfadiazine, and folic acid resulted in a complete recovery after 2 months of therapy. Although rare, acute facial nerve paralysis of unknown origin can be caused by acquired toxoplasmosis even in the immunocompetent pediatric population. Elevated titers of specific antibodies and the presence of parasite's DNA are key findings for the correct diagnosis.


Celecoxib induces apoptosis by inhibiting 3-phosphoinositide-dependent protein kinase-1 activity in the human colon cancer HT-29 cell line.
Nonsteroidal anti-inflammatory drugs, which inhibit cyclooxygenase (COX) activity, are powerful antineoplastic agents that exert their antiproliferative and proapoptotic effects on cancer cells by COX-dependent and/or COX-independent pathways. Celecoxib, a COX-2-specific inhibitor, has been shown to reduce the number of adenomatous colorectal polyps in patients with familial adenomatous polyposis. Here, we show that celecoxib induces apoptosis in the colon cancer cell line HT-29 by inhibiting the 3-phosphoinositide-dependent kinase 1 (PDK1) activity. This effect was correlated with inhibition of the phosphorylation of the PDK1 downstream substrate Akt/protein kinase B (PKB) on two regulatory sites, Thr(308) and Ser(473). However, expression of a constitutive active form of Akt/PKB (myristoylated PKB) has a low protective effect toward celecoxib-induced cell death. In contrast, overexpression of constitutive active mutant of PDK1 (PDK1(A280V)) was as potent as the pancaspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone, to impair celecoxib-induced apoptosis. By contrast, cells expressing a kinase-defective mutant of PDK1 (PDK1(K114G)) remained sensitive to celecoxib. Furthermore, in vitro measurement reveals that celecoxib was a potential inhibitor of PDK1 activity with an IC(50) = 3.5 microm. These data indicate that inhibition of PDK1 signaling is involved in the proapoptotic effect of celecoxib in HT-29 cells.


Work and cardiovascular health: viewpoint of occupational physiology.
The work-related aspects of coronary heart disease have been studied from the view-point of work physiology. The purpose of the following three studies has been to clarify how physical load at work and at leisure affects the risk of developing coronary heart disease. The first study included 120 men, mean age 41 years. They were intensively studied in the laboratory and field conditions, and were classified into four activity categories according to their work and leisure time activities. The results indicated that the highest prevalence rates of obesity, hypertension and angina pectoris symptoms were found among men doing heavy physical work and having no sporting leisure activities. The second study included a postal questionnaire to Finnish municipal employees in 1981 and 1985. Altogether 1999 women and 1419 men responded in both years. Their mean ages at the two survey times were 50.5 and 54.7 years, respectively. The 4-year incidence rates of coronary heart diseases diagnosed by the doctor (myocardial infarction, angina pectoris, coronary insufficiency, hypertension) were the highest in occupations with physical demands, both among women and especially among the men. The incidence rate of hypertension was commonest (greater than 7.0%). Among men doing physical work the incidence rate of coronary artery disease was 5.0%. The risk ratios for muscular work among men were 5.8 in the 44--49 year age group and 2.2 in the 50--58 year age group. The third project was a case-control study.(ABSTRACT TRUNCATED AT 250 WORDS)


Gut-Derived Serum Lipopolysaccharide is Associated With Enhanced Risk of Major Adverse Cardiovascular Events in Atrial Fibrillation: Effect of Adherence to Mediterranean Diet.
Gut microbiota is emerging as a novel risk factor for atherothrombosis, but the predictive role of gut-derived lipopolysaccharide (LPS) is unknown. We analyzed (1) the association between LPS and major adverse cardiovascular events (MACE) in atrial fibrillation (AF) and (2) its relationship with adherence to a Mediterranean diet (Med-diet). This was a prospective single-center study including 912 AF patients treated with vitamin K antagonists (3716 patient-years). The primary end point was a composite of MACE. Baseline serum LPS, adherence to Med-diet (n=704), and urinary excretion of 11-dehydro-thromboxane B2 (TxB2, n=852) were investigated. Mean age was 73.5 years; 42.9% were women. A total of 187 MACE (5.0% per year) occurred: 54, 59, and 74 in the first, second, and third tertile of LPS, respectively (log-rank test P=0.004). Log-LPS (hazard ratio 1.194, P=0.009), age (hazard ratio 1.083, P<0.001), and previous cerebrovascular (hazard ratio 1.634, P=0.004) and cardiac events (hazard ratio 1.822, P<0.001) were predictors of MACE. In the whole cohort, AF (versus sinus rhythm) (β 0.087, P=0.014) and low-density lipoprotein cholesterol (β 0.069, P=0.049) were associated with circulating LPS. Furthermore, Med-diet score (β -0.137, P<0.001) was predictive of log-LPS, with fruits (β -0.083, P=0.030) and legumes (β -0.120, P=0.002) negatively associated with log-LPS levels. Log-LPS and log-TxB2 were highly correlated (r=0.598, P<0.001). Log-LPS (β 0.574, P<0.001) and Med-diet score (β -0.218, P<0.001) were significantly associated with baseline urinary excretion of TxB2. In this cohort of AF patients, LPS levels were predictive of MACE and negatively affected by high adherence to Med-diet. LPS may contribute to MACE incidence in AF by increasing platelet activation.


Metabolic engineering of strains: from industrial-scale to lab-scale chemical production.
A plethora of successful metabolic engineering case studies have been published over the past several decades. Here, we highlight a collection of microbially produced chemicals using a historical framework, starting with titers ranging from industrial scale (more than 50 g/L), to medium-scale (5-50 g/L), and lab-scale (0-5 g/L). Although engineered Escherichia coli and Saccharomyces cerevisiae emerge as prominent hosts in the literature as a result of well-developed genetic engineering tools, several novel native-producing strains are gaining attention. This review catalogs the current progress of metabolic engineering towards production of compounds such as acids, alcohols, amino acids, natural organic compounds, and others.


MukB colocalizes with the oriC region and is required for organization of the two Escherichia coli chromosome arms into separate cell halves.
The circular Escherichia coli chromosome is organized by bidirectional replication into two equal left and right arms (replichores). Each arm occupies a separate cell half, with the origin of replication (oriC) at mid-cell. E. coli MukBEF belongs to the ubiquitous family of SMC protein complexes that play key roles in chromosome organization and processing. In mukBEF mutants, viability is restricted to low temperature with production of anucleate cells, reflecting chromosome segregation defects. We show that in mukB mutant cells, the two chromosome arms do not separate into distinct cell halves, but extend from pole to pole with the oriC region located at the old pole. Mutations in topA, encoding topoisomerase I, do not suppress the aberrant positioning of chromosomal loci in mukB cells, despite suppressing the temperature-sensitivity and production of anucleate cells. Furthermore, we show that MukB and the oriC region generally colocalize throughout the cell cycle, even when oriC localization is aberrant. We propose that MukBEF initiates the normal bidirectional organization of the chromosome from the oriC region.


Aging influences on working memory for hand movements: a test of the metamemory deficit hypothesis.
This study examined the role verbal labels play in working memory for hand movements between young and older adults. All young adults formed verbal labels spontaneously to remember hand movements, whereas only 20% of the older adults did so. After instructed to use verbal labels, the older adults performed at a level close to the level of the young adults. These results indicated an age-related decline of the ability to spontaneously recruit a verbal labeling strategy that can be learned and retained. However, there was no significant interaction between age group and instruction, suggesting the need for further investigation into factors other than verbal labels, including the capacity of the visuospatial memory, interference from earlier items, and fatigue.


Scaffolded reaching experiences encourage grasping activity in infants at high risk for autism.
Recent findings suggest impaired motor skill development during infancy in children later diagnosed with autism spectrum disorders (ASD). However, it remains unclear whether infants at high familial risk for ASD would benefit from early interventions targeting the motor domain. The current study investigated this issue by providing 3-month-old infants at high familial risk for ASD with training experiences aimed at facilitating independent reaching. A group of 17 high-risk (HR) infants received 2 weeks of scaffolded reaching experiences using "sticky mittens," and was compared to 72 low-risk (LR) infants experiencing the same or alternative training procedures. Results indicate that HR infants - just like LR infants - show an increase in grasping activity following "sticky mittens" training. In contrast to LR infants, evidence that motor training encouraged a preference for faces in HR infants was inconclusive.


Dietary Effect on the Proteome of the Common Octopus (Octopus vulgaris) Paralarvae.
Nowadays, the common octopus (Octopus vulgaris) culture is hampered by massive mortalities occurring during early life-cycle stages (paralarvae). Despite the causes of the high paralarvae mortality are not yet well-defined and understood, the nutritional stress caused by inadequate diets is pointed out as one of the main factors. In this study, the effects of diet on paralarvae is analyzed through a proteomic approach, to search for novel biomarkers of nutritional stress. A total of 43 proteins showing differential expression in the different conditions studied have been identified. The analysis highlights proteins related with the carbohydrate metabolism: glyceraldehyde-3-phosphate-dedydrogenase (GAPDH), triosephosphate isomerase; other ways of energetic metabolism: NADP+-specific isocitrate dehydrogenase, arginine kinase; detoxification: glutathione-S-transferase (GST); stress: heat shock proteins (HSP70); structural constituent of eye lens: S-crystallin 3; and cytoskeleton: actin, actin-beta/gamma1, beta actin. These results allow defining characteristic proteomes of paralarvae depending on the diet; as well as the use of several of these proteins as novel biomarkers to evaluate their welfare linked to nutritional stress. Notably, the changes of proteins like S-crystallin 3, arginine kinase and NAD+ specific isocitrate dehydrogenase, may be related to fed vs. starving paralarvae, particularly in the first 4 days of development.


One-sided polymerase chain reaction: the amplification of cDNA.
We report a rapid technique, based on the polymerase chain reaction (PCR), for the direct targeting, enhancement, and sequencing of previously uncharacterized cDNAs. This method is not limited to previously sequenced transcripts, since it requires only two adjacent or partially overlapping specific primers from only one side of the region to be amplified. These primers can be located anywhere within the message. The specific primers are used in conjunction with nonspecific primers targeted either to the poly(A)+ region of the message or to an enzymatically synthesized d(A) tail. Pairwise combinations of specific and general primers allow for the amplification of regions both 3' and 5' to the point of entry into the message. The amplified PCR products can be cloned, sequenced directly by genomic sequencing, or labeled for sequencing by amplifying with a radioactive primer. We illustrate the power of this approach by deriving the cDNA sequences for the skeletal muscle alpha-tropomyosins of European common frog (Rana temporaria) and zebrafish (Brachydanio rerio) using only 300 ng of a total poly(A)+ preparation. In these examples, we gained initial entry into the tropomyosin messages by using heterologous primers (to conserved regions) derived from the rat skeletal muscle alpha-tropomyosin sequence. The frog and zebrafish sequences are used in an analysis of tropomyosin evolution across the vertebrate phylogenetic spectrum. The results underscore the conservative nature of the tropomyosin molecule and support the notion of a constrained heptapeptide unit as the fundamental structural motif of tropomyosin.


Evaluation of the Family Medicine Practice in Ankara Province by Family Physicians.
Turkey's family physician or practice system was established in the beginning of the 2010 across Turkey's 81 provinces and provides low- cost health care, preventive and curative basic medical services to the population. Public health centers across Turkey have now become Family Health Centers (ASMs) as part of Turkey's efforts to harmonize its health care system with that of the European Union. The aim of This study is to analyze and evaluate the implementation and performance of Family Practice in Ankara province by family physicians. A questionnaire form of 42 question was designed and used to determine opinions of the physicians about effective service & quality improvement, patient-physician relationship, efficiency in the area of responsibility, productivity, job satisfaction and equity. The result of the study shows that family physicians were defined to be generally satisfied with the system and performance implementation and significant differences were found according to work seniority, gender and productivity of the participants. Finally this study should be taken within it's limitation. The work seniority and gender was one of the most important factor to improve satisfactions and productivity for family physicians in Turkey. The sample size was representative for the country, however, one limitation might be considered the increase of sample size in future research if appropriate funding became available in the future. This study did not have any source of funding.


Chirp imaging vibro-acoustography for removing the ultrasound standing wave artifact.
Vibro-acoustography (VA) is an imaging technique that uses the dynamic (oscillatory) radiation force of two continuous-wave (CW) ultrasound to image objects at low frequency (within the kHz range). In this technique, the dynamic radiation force is created by means of a confocused transducer emitting two ultrasound beams at slightly-shifted frequencies f1 and f2 = f1 + deltaf. It has been demonstrated previously that high-resolution images of various types of inclusions and tissues can be obtained using this technique. However, if the targeted object reflects ultrasound directly back to the transducer, standing waves are produced that result in an artifact in the VA image. The goal of this study is to remove the standing wave artifact and improve VA images by means of a new process called chirp imaging. The procedure consists of sweeping the frequencies of the primary ultrasound beams in a selected bandwidth while keeping deltaf constant during the sweep. The chirp image is produced by averaging the amplitude of the acoustic emission produced during the sweep. Vibro-acoustography chirp imaging experiments are performed on a stainless-steel sphere attached to a latex sheet in a tank of degassed water. The resulting chirp images demonstrate remarkable reduction of the standing wave artifact compared to the "fixed frequency" VA images.


The gray matter atrophy and related network changes occur in the higher cognitive region rather than the primary sensorimotor cortex after spinal cord injury.
This study used functional magnetic resonance imaging (fMRI) to explore brain structural and related network changes in patients with spinal cord injury (SCI). Thirty-one right-handed SCI patients and 31 gender- and age-matched healthy controls (HC) were included. The gray matter volume (GMV) changes in SCI patients were observed using voxel-based morphometry (VBM). Then, these altered gray matter clusters were used as the regions of interest (ROIs) for whole-brain functional connectivity (FC) analysis to detect related functional changes. The potential association between GMV and FC values with the visual analog scale (VAS), the American Spinal Injury Association (ASIA) score, and the course of injuries was investigated through partial correlation analysis. GMV of the frontal, temporal, and insular cortices was lower in the SCI group than in the HC group. No GMV changes were found in the primary sensorimotor area in the SCI group. Besides, the altered FC regions were not in the primary sensorimotor area but in the cingulate gyrus, supplementary motor area, precuneus, frontal lobe, and insular. Additionally, some of these altered GMV and FC regions were correlated with ASIA motor scores, indicating that higher cognitive regions can affect motor function in SCI patients. This study demonstrated that gray matter and related network reorganization in patients with SCI occurred in higher cognitive regions. Future rehabilitation strategies should focus more on cognitive functions.


Quantitative DNA methylation analysis based on four-dye trace data from direct sequencing of PCR amplificates.
Methylation of cytosines in DNA plays an important role in the regulation of gene expression, and the analysis of methylation patterns is fundamental for the understanding of cell differentiation, aging processes, diseases and cancer development. Such analysis has been limited, because technologies for detailed and efficient high-throughput studies have not been available. We have developed a novel quantitative methylation analysis algorithm and workflow based on direct DNA sequencing of PCR products from bisulfite-treated DNA with high-throughput sequencing machines. This technology is a prerequisite for success of the Human Epigenome Project, the first large genome-wide sequencing study for DNA methylation in many different tissues. Methylation in tissue samples which are compositions of different cells is a quantitative information represented by cytosine/thymine proportions after bisulfite conversion of unmethylated cytosines to uracil and PCR. Calculation of quantitative methylation information from base proportions represented by different dye signals in four-dye sequencing trace files needs a specific algorithm handling imbalanced and overscaled signals, incomplete conversion, quality problems and basecaller artifacts. The algorithm we developed has several key properties: it analyzes trace files from PCR products of bisulfite-treated DNA sequenced directly on ABI machines; it yields quantitative methylation measurements for individual cytosine positions after alignment with genomic reference sequences, signal normalization and estimation of effectiveness of bisulfite treatment; it works in a fully automated pipeline including data quality monitoring; it is efficient and avoids the usual cost of multiple sequencing runs on subclones to estimate DNA methylation. The power of our new algorithm is demonstrated with data from two test systems based on mixtures with known base compositions and defined methylation. In addition, the applicability is proven by identifying CpGs that are differentially methylated in real tissue samples.