Targeting Lewis X oligosaccharide-modified liposomes encapsulated with house dust mite allergen Der f 2 to dendritic cells inhibits Th2 immune response.
Allergen-specific immunotherapy (AIT) is the only curative treatment for allergic diseases. However, the long desensitization phase and potentially dangerous allergic side effects limit its broad application. Therefore, safer and more effective vaccines are required. Targeting dendritic cells (DCs) with novel allergen conjugates is a promising strategy for AIT. In this study, a novel vaccine with a DC-targeting effect for AIT was constructed. Liposomes were used as vehicles, and a targeted nanovaccine (Lex-lip-Der f 2) was constructed by loading the recombinant group 2 allergen of Dermatophagoides farinae (Der f 2) and conjugating with the DC-SIGN ligand Lewis X. The effect of the vaccine on DCs and T cell responses and the safety of the vaccine were investigated in vitro. The results showed that the Lex-lip-Der f 2 vaccine was spherical, with size of approximately 128 nm. The protein-loading capacity of the vaccine was 0.106 ± 0.001 mg per mg liposome and protein was gradually released from the liposomes during the first 12 h. Lex-lip-Der f 2 was taken up more efficiently by DCs than non-targeted liposomes or free Der f 2. Besides, Lex-lip-Der f 2 significantly inhibited the release of IL-4, IL-6, and TNF-a from DCs. Accordingly, Der f 2-lip loaded DCs significantly decreased IL-4 levels in autologous naïve CD4+T cells. Moreover, Lex-lip-Der f 2-treated basophils showed lower activation levels. These results suggest that DC-SIGN targeting mediated by Lewis X could inhibit the Th2 cell response and improve vaccine safety, and may be a novel vaccination strategy.


Hyperpolarized 129Xe Pulmonary MRI and Asymptomatic Atrial Septal Defect.
In an asymptomatic 19-year-old who regularly underwent cardiopulmonary fitness testing for national lifeguard-accreditation, 129Xe MRI unexpectedly revealed an abnormally augmented RBC signal and RBC-to-alveolar-capillary-tissue ratio with spatially homogeneous ventilation, tissue barrier, and RBC images. Pulmonary function was normal, but cardiopulmonary follow-up including transthoracic and transesophageal echocardiogram, heart catheterization, and contrast-enhanced cardiac CT imaging led to the diagnosis of a large (20 × 27 mm) secundum atrial septal defect (ASD) with a net right-to-left shunt (Qp:Qs = 0.5) and normal pulmonary pressures. This novel, unexpected case revealed that 129Xe RBC signal intensity likely reflected erythrocytosis, compensatory to the abnormal cardiovascular hemodynamics that resulted from a large congenital ASD. Unlike ASD cases that present with dyspnea and exercise limitation, this 129Xe MRI abnormality was detected in an asymptomatic teenager. This is the first report of asymptomatic adult congenital heart disease diagnosed subsequent to novel 129Xe MRI that led to early intervention, avoiding long-term complications of cyanosis, including ventricular fibrosis and thromboembolic and bleeding risks.


Scanning electron microscopic evaluation of the surface characteristics of 4 microkeratome systems in human corneas.
To evaluate the surface characteristics of keratectomy specimens created with 4 microkeratomes by scanning electron microscopy (SEM). Two university institutions. Keratectomies were performed in 10 fresh, unfrozen human eyes with 1 of 4 microkeratomes: the Draeger rotary microkeratome (Storz Instruments), the Universalkeratome (Phoenix), the Automated Corneal Shaper (Bausch & Lomb), or the Hansatome (Bausch & Lomb). The beds and edges of the keratectomy specimens were then examined by SEM. Standardized photographs were taken, and the surface and cut-edge characteristics of the keratectomies were compared. Irregularities were noted in the cut edges of Bowman's membrane and the surface of the keratectomy beds with all instruments. The irregularities were most prominent in corneas cut with the Draeger instrument and least prominent in those cut with the Universalkeratome. The serrations in Bowman's membrane and the irregularities in the keratectomy bed appeared to be related to the blade movement in oscillating blade instruments. All 4 microkeratomes caused some degree of surface irregularity. The Draeger microkeratome, which uses a continuously rotating blade, resulted in the greatest number of Bowman's membrane serrations and irregularities in the keratectomy bed. The tissue irregularities may be related to the development of pressure ridges ahead of the blade. In instruments that use an oscillating blade, chatter marks appear to be related to the reciprocating movement of the blade as it travels across the corneal stroma.


Benign uterine leiomyoma suspected of sarcomatous change on an ultrasound scan and computerized tomography.
We report a 54-year-old postmenopausal woman with a leiomyoma which on an ultrasound scan and on computed tomography (CT) had cystic areas and extensive necrosis suggesting sarcomatous change.


Estimating population size and demographic characteristics of lesbian, gay, bisexual, and transgender youth in middle school.
To estimate the size and demographic characteristics of the lesbian, gay, bisexual, and transgender youth populations using data from the 2011 Youth Risk Behavior Survey (YRBS) administered in San Francisco Unified School District middle schools. The YRBS was administered to a stratified random sample of 2,730 youth (grades 6-8) across all 22 public middle schools in San Francisco. Cross-tabulations using complex samples analyses were used to derive population estimates and confidence intervals. Results show that 3.8% of middle school students identify as lesbian, gay, or bisexual, and 1.3% of middle school students identify as transgender. To improve our understanding of the size of these populations across the nation, researchers conclude it is imperative that all YRBS administration sites include items on sexual orientation and gender identity as they would any other demographic item, such as race/ethnicity, sex, or age. The current lack of reliable data on the size and characteristics of the lesbian, gay, bisexual, and transgender youth population limits the capacity of policy makers, administrators, and practitioners to address their needs.


Low plasma vasopressin/norepinephrine ratio predicts septic shock.
To evaluate if low plasma vasopressin and high norepinephrine concentrations predict grave prognosis of sepsis, a prospective sample of consecutive patients visiting the emergency department of a university teaching hospital who met the American College of Chest Physicians criteria of sepsis or severe sepsis was enrolled. Besides septic workup, we measured serum vasopressin and norepinephrine concentrations to correlate the impending outcome. One hundred eighty-two patients aged 27 to 99 years met the inclusive criteria and were classified as those with septic shock (n = 72), severe sepsis (n = 56), and those with sepsis only (n = 54) according to the outcome within 6 hours. Thirty healthy subjects were included as control. The plasma vasopressin level at baseline was significantly lower for those who finally developed septic shock (septic shock group, 3.6 +/- 2.5 pg/mL; 95% confidence interval [CI], 3.0-4.2 pg/mL; severe sepsis group, 21.8 +/- 4.1 pg/mL, 95% CI, 20.7-22.9 pg/mL; sepsis group, 10.6 +/- 6.5 pg/mL, 95% CI, 8.8-12.4 pg/mL, P < .001), whereas the norepinephrine level was highest for the same group (septic shock group, 3650 +/- 980 pg/mL, 95% CI, 3420-3880 pg/mL; severe sepsis group, 3600 +/- 1000 pg/mL, 95% CI, 3330-3870 pg/mL; sepsis group, 1720 +/- 320 pg/mL, 95% CI, 1630-1810 pg/mL). The vasopressin/norepinephrine ratio was significantly lower for the patients with final diagnosis of septic shock (P < .001). The mean interval between the time of samples drawn and the time of the most severe occurring sequelae was 2.4 +/- 0.8 hours. Receiver operating characteristic analysis revealed that the vasopressin/norepinephrine ratio 1 x 10(-3) had a sensitivity of 97% (95% CI, 90%-99%) and a specificity of 85% (95% CI, 78%-91%) for detecting impending septic shock. Low serum vasopressin/norepinephrine ratio can predict impending septic shock.


New nanostructured extracellular potassium ion probe for assay of cellular K+ transport.
The concentration of potassium ion is an important indicator for human health, and its abnormality is often accompanied by various diseases. However, most tools currently used to study potassium ion transport are low throughput. Herein, we reported a new K+ fluorescent nanoprobe CP1-KS with high selectivity and sensitivity to K+ (fluorescence enhanced factor was up to 9.91 at 20 mM K+). The polymeric fluorescent probe CP1-KS was composed of the small-molecular K+ indicator KS and amphiphilic copolymer CP1. This sensor can be easily and uniformly dispersed in cell culture medium and is suitable for high throughput analysis. To assess the utility of the probe CP1-KS in biological field, this probe was employed as an extracellular fluorescent probe to monitor the efflux of K+ from cells (E coli, B. Subtilis 168, Hela and MCF-7 cells) under various stimulation including lysozyme, nigericin, digitonin, and ATP. Results demonstrated that CP1-KS is an effective analysis tool for extracellular K+ concentration. We believe that the nanoprobe has great potential in antibacterial drug screening, K+ ionophore function, K+ channel activity, cell membrane permeability analysis or other K+ related field in the future.


Analytical methods for thorium determination: a journey from conventional methods to novel applications.
Thorium may be determined using a variety of analytical methods. These may be based on chemical or physical principles or may make use of the radioactive decay of the individual thorium isotopes. The method to be used for a certain analytical purpose is selected as a function of the matrix of the sample and of the concentration of thorium. The present paper describes different methodologies, their range of application and discusses selected results. The methods described cover a concentration range of more than twelve orders of magnitude, i.e. from concentrated solutions to micrometre-sized particles. Emphasis is given to active radiometric techniques, chemical and instrumental analysis.


The current preferred female lip ratio.
Perception of beauty is influenced by the individual's demographic background and characteristics. However, objective measurements and ratios remain the foundation for aesthetic evaluations. This study aimed to elucidate if there exists a universally applicable ideal upper to lower lip ratio. An interactive online survey was designed. Modifiable ranges of lip ratios were achieved via digital alteration, enabling participants to change the ratio of a single female model's lips. The questionnaire was translated into multiple languages and sent to more than 9000 plastic surgeons and the general public worldwide. Demographic data were collected and analysis of variance was used to investigate respective lip ratio preferences. A total of 1011 responses from 35 different countries (response rate of 14%) was gathered. The majority of survey takers (60%) chose the 1.0:1.0 lip ratio as most attractive. No differences were found in respect to lip ratio preference and the self-reported ethnicity. However, interesting preferences prevailed when analyzing the subgroups regarding lower lip size. Age, gender, country of residence, and profession significantly impact individual upper to lower lip ratio preferences. However, a 1.0:1.0 lip ratio can apparently be considered most pleasing in females.


Synthesis and properties of O6-methyldeoxyguanylic acid and its copolymers with deoxycytidylic acid.
This paper describes the synthesis of O6-methyldeoxyguanosine triphosphate (m6dGTP) and its copolymerization to high molecular weight polymer with deoxycytidylic acid. The monomer, m6dGTP, was synthesized from deoxyguanosine first protected by acetylation of the sugar hydroxyls, and then chlorinated in the 6-position with POCl3. The product, 6-chloro-3',5'-di-O-acetyl deoxyguanosine, was converted to O6-methyldeoxyguanosine with sodium methoxide and phosphorylated in the 5' position with carrot phosphotransferase. Monophosphate was converted chemically to the triphosphate and copolymerized with dCTP by terminal deoxynucleotidyl transferase. The resulting template, which contained O6-methylguanine, was tested for its ability to direct RNA synthesis by bacterial RNA polymerase. The presence of O6-methylguanine was shown to lead to the misincorporation of UMP in the product polymer, thus strengthening the hypothesis that O6-methylguanine is a promutagenic base.


Breakthrough Cancer Treatments Raise Difficult Questions.
The approval of a first-of-its kind cancer therapy creates new challenges related to safety, access, and costs.


L-Asparaginase delivery systems targeted to minimize its side-effects.
L-asparaginase (L-ASP) is one of the key enzymes used in therapeutic applications, particularly to treat Acute Lymphocytic Leukemia (ALL). L-asparagine is a non-essential amino acid, which means that it can be synthesized by the body and is not required to be obtained through the diet. The synthesis of L-asparagine occurs primarily in the liver, but it also takes place in other tissues throughout the body. In contrast, leukemic cells cannot synthesize L-asparagine due the absence of L-asparagine synthetase and should obtain it from circulating sources for protein synthesis and cell division processes to ensure their vital functions. L-ASP catalyzes the deamination process of L-asparagine amino-acid into aspartic acid and ammonia, depriving leukemic cells of asparagine. This leads to decreased protein synthesis and cell division in tumor cells. However, using L-ASP has side effects, such as hypersensitivity or allergic reaction, antigenicity, short half-life, temporary blood clearance, and toxicity. L-ASP immobilization can minimize the side effects of L-ASP by stopping the immune system from attacking non-human enzymes and improving the enzyme's performance. The first strategy includes modification of enzyme structure, such as covalent binding (conjugation), adsorption to the support material and cross-linking of the enzyme. The chemical modification of residues, often nonspecific, changes the enzyme's hydrophobicity and surface charge, lowering the enzyme's activity. Also, the first strategy exposes the enzyme's surface to the environment. This eliminates its performance and does not allow targeted delivery of the enzyme. The second strategy is based on the entrapment of the enzyme inside the protecting structure or encapsulation. This strategy offers the same benefits as the first. Still, it also enables reducing toxicity, prolonging in vivo half-life, enhancing stability and activity, enables a targeted delivery and controlled release of the enzyme. Compared to the first strategy, encapsulation does not modify the chemical structure of the enzyme since L-ASP is only effective against leukemia in its native tetrameric form. This review aims to present state of the art in L-ASP formulations developed for reducing the side effects of L-ASP, focusing on describing improvements in their safety. The primary focus in the field remains to be improving the overall performance of the L-ASP formulations. Almost all encapsulation systems allow reducing immune response due to screening the enzyme from antibodies and prolonging its half-life. However, the enzyme's activity and stability depend on the encapsulation system type. Therefore, the selection of the right encapsulation system is crucial in therapy due to its effect on the performance parameters of the L-ASP. Biodegradable and biocompatible materials, such as chitosan, alginate and liposomes, mainly attract the researcher's interest in enzyme encapsulation. The research trends are also moving towards developing formulations with targeted delivery and increased selectivity.


The reaction of tryptophan with cystine during acid hydrolysis of proteins. Formation of tryptathionine as a transient intermediate in a model system.
Under the conditions commonly used to hydrolyze proteins with 6 M HCl, tryptophan reacted with cystine to give a transient intermediate, which was isolated and identified as 2-(2-amino-2-carboxyethylthio)tryptophan (tryptathionine) by NMR studies, etc. Studies on the formation and degradation of the above compound showed that beta-3-oxoindolylalanine and cysteine, which were previously reported to be the main degradation products of tryptophan and cystine, respectively, are formed by the hydrolysis of this intermediate during the course of the reaction.


Triphenyltin impairs a protein kinase A (PKA)-dependent increase of cytosolic Na+ and Ca2+ and PKA-independent increase of cytosolic Ca2+ associated with insulin secretion in hamster pancreatic beta-cells.
Oral administration of triphenyltin chloride (TPT) (60 mg/kg body weight) inhibits the insulin secretion by decreasing the cytoplasmic Ca2+ concentration ([Ca2+]i) induced by glucose-dependent insulinotropic polypeptide (GIP) in pancreatic beta-cells of the hamster. To test the possibility that the abnormal level of [Ca2+]i induced by TPT administration could be due to a defect in the cAMP-dependent cytoplasmic Na+ concentration ([Na+]i) in the beta-cells, we investigated the effects of TPT administration on the changes of [Na+]i induced by GIP, glucagon-like peptide-1 (GLP-1), or forskolin, an activator of adenylyl cyclase, and on the changes of [Na+]i or [Ca2+]i induced by 6-Bnz-cAMP, an activator of protein kinase A (PKA), and 8-pCPT-2'-O-Me-cAMP, an activator of Epac. The [Na+]i and [Ca2+]i were measured in islet cells loaded with sodium-binding benzofuran isophthalate (SBFI) and fura-2, respectively. In the presence of 135 mM Na+, TPT administration significantly reduced the rise in [Na+]i by 10 nM GLP-1, 10 microM forskolin, and 50 microM 6-Bnz-cAMP, but had not effect in a Na+-free medium. In the presence of 135 mM Na+, TPT administration also reduced the rise in [Ca2+]i by 8-pCPT-2'-O-Me-cAMP plus10 microM H-89, a inhibitor of PKA, and 6-Bnz-cAMP. Moreover, TPT administration significantly reduced the insulin secretion by 2 mM db-cAMP, GLP-1, GIP, and 8-pCPT-2'-O-Me-cAMP with and without H-89, and that by 6-Bnz-cAMP and forskolin. Our study suggested that TPT has inhibitory effects on the cellular Ca2+ response due to a reduced Na+ permeability through PKA-dependent mechanisms in hamster islet cells. Also TPT has the reduction of [Ca2+]i related to Na+-dependent insulin secretion after an activation of Epac.


Video fluorescein angiography: method and clinical application.
Video fluorescein angiography, combined with a picture analyzing system, is a clinically applicable, objective method of evaluating the retinal blood-flow parameters. Optical density measurements were performed on videorecordings of fluorescence angiograms by means of a picture-analyzing system in order to determine the circulation parameters of the retina. These included: the arm-retina time (ART), the arteriovenous passage time (AVP), and the mean arterial dye-bolus velocity (MDV). Normal values for these parameters were derived from measurements in 75 healthy volunteers. The mean arm-retina time (ART) was 11.2 +/- 3.3 s, the mean arteriovenous passage time (AVP) 1.45 +/- 0.4 s and the mean arterial dye-bolus velocity (MDV) 6.39 +/- 1.7 mm/s. No significant correlation could be shown between pulse or blood pressure and one of the retinal circulation parameters. A group of ten healthy volunteers was examined twice in order to obtain the intraindividual variation for the measuring parameters. The coefficient of variation for the ART was 18%, 10% for the AVP, and 26% for the MDV.


Biomarker-Driven Studies With Multi-targets and Multi-drugs by Next-Generation Sequencing for Patients With Non-Small-Cell Lung Cancer: An Open-Label, Multi-center, Phase II Adaptive Umbrella Trial and a Real-World Observational Study (CTONG1702&CTONG1705).
With rare genetic variations having been increasingly recognized at a preclinical stage, a variety of early-phase clinical trials have been launched. Due to the low incidence rate of these variations, although the sample size of trials are small, it still needs a large number of patients for screening. With the advent of next-generation sequencing (NGS), multiple genetic variations can be detected simultaneously. Multiple biomarkers and agents can be evaluated using umbrella clinical trials, which rapidly and effectively screen and enroll patients for parallel sub-studies using NGS. We designed an open-label, multi-center, phase II clinical trial CTONG1702. This is an adaptive umbrella trial that will evaluate the efficacy and safety of several biomarker-driven agents, including tyrosine kinase inhibitors (TKIs) and a PD-1 inhibitor, in stage IIIB to IV patients (eighth AJCC) with non-small-cell lung cancer (NSCLC) patients. Patients will be enrolled in parallel sub-studies based on the results of NGS and PD-L1 IHC analysis. Patients who are not eligible for CTONG1702 will be enrolled in the observational real-world study CTONG1705. This study aims to develop a large-scale genomic database and explore the relationship between genetic variations in NSCLC patients and clinical outcomes. The adaptive umbrella trial will evaluate multi-targets and multi-drugs in advanced NSCLC patients (CTONG1702). In addition, the simultaneously initiated real-world study will provide additional data for clinical practice (CTONG1705).


What makes gambling news?
This paper examines print media coverage of casino and electronic gambling in one Canadian province from 1992 to 1997. It provides a theme analysis of content of 234 gambling stories printed in the top two daily newspapers in Nova Scotia. The findings of our content analysis indicate that pro-gambling corporate and political newspaper sources waged a successful media campaign and constructed a powerful public rhetoric in support of new gambling products, services, and institutions. The media, for their part, gave visibility and form to these structured messages. They helped create expectations about gambling and economics and gambling and government. Law and order, and moral and medical discourses about gambling, we discovered, were minor representations in the news coverage, although moral narratives were a pervasive secondary theme in much of the reporting. At bottom, the press produced a "politics of truth" about gambling that was both an external exercise of power and an internal organizational production.


The effect of local species composition on the distribution of an avian invader.
Estimating the potential distribution of invasive species has been primarily achieved by employing species distribution models (SDM). Recently introduced joint species distribution models (JSDM) that include species interactions are expected to improve model output. Here we compare the predictive ability of SDM and JSDM by modelling the distribution of one of the most prolific avian invaders in the world, the common myna (Acridotheres tristis), in a recent introduction in Israel. Our results indicate that including information on the local species composition did not improve model accuracy, possibly because of the unique characteristics of this species that include broad environmental tolerance and behavior flexibility. However, the JSDM provided insights into co-occurrence patterns of common mynas and their local heterospecifics, suggesting that at this time point, there is no evidence of species exclusion by common mynas. Our findings suggest that the invasion potential of common mynas depends greatly on urbanization and less so on the local species composition and reflect the major role of anthropogenic impact in increasing the distribution of avian invaders.


Practical uncertainty reduction and quantification in shock physics measurements.
We report the development of a simple error analysis sampling method for identifying intersections and inflection points to reduce total uncertainty in experimental data. This technique was used to reduce uncertainties in sound speed measurements by 80% over conventional methods. Here, we focused on its impact on a previously published set of Mo sound speed data and possible implications for phase transition and geophysical studies. However, this technique's application can be extended to a wide range of experimental data.


Color stability and hardness in dental composites after accelerated aging.
To investigate the color and microhardness changes of five chemically- and five light-curing composites as a function of accelerated aging from light exposure. From each material five composite specimens were embedded in epoxy resin prior to determining the Knoop microhardness of the surface. For analyzing the color DeltaE*=f((L*a*b*)) with a spectrophotometer, three discs per composite were prepared. After measuring the baseline for hardness and color the same specimens were exposed to a xenon arc light and water in a Weather-Ometer machine for a total radiant energy of 150 kJ/m2 and 122 h. The microhardness and the color were again determined following the aging treatment. Each material showed a significant increase in hardness after aging treatment (p<0.05). Comparing the hardness changes (in %) of the light-cured materials with the chemically cured materials, no significant difference could be found. Perceptible color differences could be observed for all the materials. Three brands showed small differences with DeltaE*=1.6-2.2, while four composites had DeltaE* ranging from 6.2 to 15.5. A significant correlation between hardness values and color changes could not be established. The findings suggest that, since light-curable materials showed significantly more resistance to color changes after accelerated aging by light and water than chemically-cured materials, they may be more esthetically acceptable. Color changes were not correlated with surface hardness changes of the materials after aging.


Effect from low-level exposure of oxytetracycline on abundance of tetracycline resistance genes in arable soils.
To evaluate the effect from low-level exposure of antibiotics on the abundance of antibiotic resistance genes, unpolluted arable soils were treated with oxytetracycline (OTC)-containing manure, with OTC (0, 2, 20, or 70 μg kg(-1)) added every 2 weeks on 10 occasions. Six tetracycline resistance genes [TRGs-tet(A), tet(L), tet(M), tet(Q), tet(O), and tet(W)] and the 16S ribosomal RNA (rRNA) gene were monitored using real-time quantitative polymerase chain reaction. The relative abundance of tet(A), tet(L), tet(M), and tet(Q) genes in soil increased 10-1000 times after application of OTC-containing manure. Tet(A) abundance per unit of residual OTC on day 140 was 1.53-4.42 times higher than that on day 28, while tet(L) abundance was 1.04-1.74 times higher. Treatment with >40 μg kg(-1) OTC significantly increased abundance of tet(A) and tet(L), while tet(M) and tet(Q) abundance was positively correlated (R (2) = 0.965 and 0.932, p < 0.01) with residual OTC concentrations. There was a significant accumulation of TRGs associated with low-level OTC exposure in arable soils. Besides OTC residual, the effects from exposure time and application frequencies should also be considered to limit the increase in abundance of tet(A) and tet(L).


Mapping allele with resolved carrier status of Robertsonian and reciprocal translocation in human preimplantation embryos.
Reciprocal translocations (RecT) and Robertsonian translocations (RobT) are among the most common chromosomal abnormalities that cause infertility and birth defects. Preimplantation genetic testing for aneuploidy using comprehensive chromosome screening for in vitro fertilization enables embryo selection with balanced chromosomal ploidy; however, it is normally unable to determine whether an embryo is a translocation carrier. Here we report a method named "Mapping Allele with Resolved Carrier Status" (MaReCs), which enables chromosomal ploidy screening and resolution of the translocation carrier status of the same embryo. We performed MaReCs on 108 embryos, of which 96 were from 13 RecT carriers and 12 were from three RobT carriers. Thirteen of the sixteen patients had at least one diploid embryo. We have confirmed the accuracy of our carrier status determination in amniotic fluid karyotyping of seven cases as well as in the live birth we have thus far. Therefore, MaReCs accurately enables the selection of translocation-free embryos from patients carrying chromosomal translocations. We expect MaReCs will help reduce the propagation of RecT/RobT in the human population.


Comparison of Lateral and Dorsal Recumbency during Endoscope-Assisted Oophorectomy in Mature Pond Sliders (Trachemys scripta).
Ovariectomy via the prefemoral fossa is a well-described technique for sterilization in chelonians. The choice between lateral and dorsal recumbency is generally left to the surgeon's preference, with no data supporting an objective superiority of one over the other. Twenty-four sexually mature female pond sliders (Trachemys scripta) were enrolled for elective prefemoral endoscope-assisted oophorectomy, and were randomly divided in two groups: 12 animals were placed in right lateral recumbency with a left fossa approach (Group A), and 12 in dorsal recumbency with a right fossa approach (Group B). Scoring systems were applied to assess the ease of access to the coelomic cavity, and the ease of identification of the ovary opposite to the surgical incision. A negative correlation was found between the body weight of the animals and the ease of access to the coelom (p = 0.013), making the access easier in smaller animals. No significant difference was found between the two groups in terms of ease of access to the coelomic cavity, first ovary removal time, ease of identification of the second ovary, second ovary removal time, or total surgery time.


Pulmonary complications in dogs with acute presentation of pancreatitis.
In humans, respiratory complications in patients with acute pancreatitis (AP) are a common life-threatening comorbidity. Since possible lung impairment has not been individually evaluated in canine AP, the aims of the present study were to: (1) describe the prevalence, types and severity of pulmonary complications in dogs with acute presentation of AP, and (2) evaluate their association with mortality. AP diagnosis was based on compatible clinical and laboratory parameters, abnormal canine pancreatic-lipase test, and positive abdominal ultrasound within 48 h from admission. The canine acute pancreatitis severity score (CAPS) was calculated for each dog at admission. Arterial blood gas analysis and thoracic radiography were performed at admission. Thoracic radiography was classified on the basis of pulmonary pattern (normal, interstitial or alveolar) and a modified lung injury score (mLIS) was applied to the ventrodorsal projections for each dog. VetALI/VetARDS were diagnosed using current veterinary consensus. Dogs were divided into non-survivors or survivors (hospital discharge). Clinical, radiological and blood gas parameters collected at presentation were compared between survivors and non-survivors and associated with mortality. This prospective cohort study included twenty-six client-owned dogs with AP. Twelve out of twenty-six dogs (46%) died or were euthanized. At admission, thirteen dogs showed respiratory distress at physical examination, which was associated with death (P < 0.001). Radiographic abnormalities were found in twenty-one dogs: alveolar (n = 11) and interstitial pattern (n = 10). Radiographic alterations and mLIS score were both associated with death (P = 0.02 and P = 0.0023). The results of the arterial blood-gas evaluation showed that non-survivors had lower PaCO2 and HCO3- levels, and higher A-a gradient than survivors (P = 0.0014, P = 0.019 and P = 0.004, respectively). Specifically, three dogs had aspiration pneumonia, and VetALI was diagnosed in nine dogs (34.6%), and no dogs met the criteria for VetARDS. The presence of VetALI was associated with mortality (P < 0.001). As with humans, possible lung impairments, such as VetALI, should be investigated in dogs with acute presentation of pancreatitis.


[Recommendations for the diagnosis and treatment of persistent postsurgical symptoms in Hirschsprung disease].
Hirschsprung disease is characterized by the lack of migration of intrinsic parasympathetic ganglia from neural crest and consequently absence of them at varying length of the bowel, resulting in functional obstruction. The incidence is 1 per 5000 births. After surgery, short term and long term comorbidity commonly occurs. The aim of this article is to revise the main causes of ongoing symptoms after surgery in Hirschsprung disease patients and to show a diagnostic and therapeutic algorithm that can be developed in our community.


Metal ion requirement of bifunctional UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase from rat liver.
The metal ion requirement for both enzymatic activities of the bifunctional UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (E.C. 188.8.131.52/18.104.22.168), the key enzyme of N-acetylneuraminic acid biosynthesis in rat liver, was investigated. UDP-N-acetylglucosamine 2-epimerase was active in imidazole/HCl buffer in the complete absence of any metal ion. 200 mM Na+, K+, Rb+ and Cs+ activated enzyme activity up to five-fold, whereas lower concentrations of these monovalent metal ions showed only a small effect on UDP-N-acetylglucosamine 2-epimerase activity. In sodium phosphate buffer the enzyme activity was increased by 0.5 mM Mg2+, Sr2+, Ba2+ and Mn2+, while in the presence of 200 mM NaCl UDP-N-acetylglucosamine 2-epimerase activity showed a stronger activation by these divalent metal ions. In imidazole/HCl buffer, UDP-N-acetylglucosamine 2-epimerase activity was partially inhibited by 0.5 mM Be2+, Mg2+, Ba2+, Mn2+, Sn2+ and Fe2+, and completely inhibited by 0.5 mM Zn2+ and Cd2+. Divalent metal ions were essential for N-acetylmannosamine kinase activity, the most effective being Mg2+, followed by Mn2+ and Co2+. The optimal concentration of these metal ions was 3 mM. Less effective were Ni2+ and Cd2+, whereas Ca2+, Ba2+, Cu2+, Fe2+ and Zn2+ showed no effect on enzyme activity.


Bartonella-host-cell interactions and vascular tumour formation.
Bartonellae are arthropod-borne bacterial pathogens that typically cause persistent infection of erythrocytes and endothelial cells in their mammalian hosts. In human infection, these host-cell interactions result in a broad range of clinical manifestations. Most remarkably, bartonellae can trigger massive proliferation of endothelial cells, leading to vascular tumour formation. The recent availability of infection models and bacterial molecular genetic techniques has fostered research on the pathogenesis of the bartonellae and has advanced our understanding of the virulence mechanisms that underlie the host-cell tropism, the subversion of host-cell functions during bacterial persistence, as well as the formation of vascular tumours by these intriguing pathogens.


Effect of protein kinase C on transmembrane calcium fluxes in HaCaT keratinocytes.
Capacitive calcium influx is associated with the release of calcium from internal stores and participates in intracellular calcium homeostasis. In keratinocytes, its activation is linked to the stimulation of the phospho-inositide (PI) pathway and seems to be altered in psoriasis. An overnight treatment of isolated HaCaT keratinocytes with phorbol 12-myristate 13-acetate (PMA) selectively downregulated the classical, calcium-dependent protein kinase C (PKC) isoenzyme PKC alpha in preconfluent cells. This was parallelled by an increased capacitative calcium influx with no effects on the PI pathway. These observations were strengthened in measurements using cyclopiazonic acid which revealed a 47% increase in PMA pretreated as compared with control cells in the calcium influx rate through store-operated calcium channels (SOC-s) following the emptying of the intracellular calcium stores. In confluent as compared with preconfluent cultures PKC epsilon was markedly increased, while other isoenzymes were not affected. In parallel, the kinetics of capacitative calcium influx were altered, showing clear inactivation. PMA pretreatment in these cells had little effect on PKC alpha but downregulated both PKC beta and PKC epsilon, and did not increase the influx through SOC-s. These observations support the differential regulation of SOC-s by PKC and suggest the involvement of several PKC isoenzymes in human keratinocytes.


Characterization and heterologous expression of plasmalogen synthase MeHAD from Megasphaera elsdenii.
Plasmalogens (Pls) are vinyl-ether bond-containing glycerophospholipids or glycosyl diradyl glycerols, and are of great importance in the physiological functions and stability of cell membrane. Here, we identified and characterized that the plasmalogen synthase MeHAD from anaerobic Megasphaera elsdenii was responsible for vinyl-ether bond formation. Different from the 2-hydroxyacyl-CoA dehydratase (HAD) family plasmalogen synthase PlsA-PlsR which are encoded by two genes in Clostridium perfringens, the HAD homolog (MeHAD) encoded by a single gene MELS_0169 was found in M. elsdenii. By heterologous expression of the MeHAD gene into a nonplasmalogen-producing Escherichia coli strain, the expressed MeHAD was found to be located in the cell membrane region. Plasmalogens were detected in the recombinant strain using GC-MS and LC-MS, demonstrating that MeHAD was the key enzyme for plasmalogen synthesis. Moreover, the synthesized plasmalogens could enhance the oxidative stress-resistance and osmotic pressure-resistance of the recombinant strain, probably due to the ROS scavenging and decreased membrane permeability by the plasmalogens, respectively. The four-cysteine (Cys125, Cys164, Cys445 and Cys484) site-mutant of MeHAD, which were predicted binding to the [4Fe-4S] cluster, was unable to synthesize plasmalogens, indicating that the cysteines are important for the catalytic activity of MeHAD. Our results revealed the single gene encoded plasmalogen synthase in M. elsdenii and established a recombinant E. coli strain with plasmalogen production potential.


Alcohol, drug misuse and suicide attempts: unrecognised causes of out of hospital cardiac arrests admitted to intensive care units.
To assess the contribution of alcohol, drug abuse and suicide attempts to out of hospital cardiac arrests (OHCA) who are admitted to our intensive care unit (ICU). Retrospective review of all OHCA admitted to the ICU over a 2-year period. There were 26 OHCA. Six patients survived, all of whom had a cardiac aetiology for their arrest. Ten patients arrested due to external factors (drug misuse n = 4, alcohol excess n = 1, suicide attempts n = 4 and accidental choking n = 1). All of the patients who arrested secondary to external factors were young (37.2 +/- 13.58 years), 90% were male and all died in hospital. All of the cases of drug misuse involved cocaine. Alcohol, drug misuse and suicide attempts contribute significantly to the number of OHCA which are admitted to ICU. Moreover, cocaine usage has contributed to a number of OHCA in our study.


Optimization of Deep Learning Based Brain Extraction in MRI for Low Resource Environments.
Brain extraction is an indispensable step in neuro-imaging with a direct impact on downstream analyses. Most such methods have been developed for non-pathologically affected brains, and hence tend to suffer in performance when applied on brains with pathologies, e.g., gliomas, multiple sclerosis, traumatic brain injuries. Deep Learning (DL) methodologies for healthcare have shown promising results, but their clinical translation has been limited, primarily due to these methods suffering from i) high computational cost, and ii) specific hardware requirements, e.g., DL acceleration cards. In this study, we explore the potential of mathematical optimizations, towards making DL methods amenable to application in low resource environments. We focus on both the qualitative and quantitative evaluation of such optimizations on an existing DL brain extraction method, designed for pathologically-affected brains and agnostic to the input modality. We conduct direct optimizations and quantization of the trained model (i.e., prior to inference on new data). Our results yield substantial gains, in terms of speedup, latency, through-put, and reduction in memory usage, while the segmentation performance of the initial and the optimized models remains stable, i.e., as quantified by both the Dice Similarity Coefficient and the Hausdorff Distance. These findings support post-training optimizations as a promising approach for enabling the execution of advanced DL methodologies on plain commercial-grade CPUs, and hence contributing to their translation in limited- and low- resource clinical environments.


Effect of alumina (Al2O3) nanoparticles and macroparticles on Trigonella foenum-graceum L. in vitro cultures: assessment of growth parameters and oxidative stress-related responses.
The impact of 100 μg ml-1 alumina (Al2O3) nanoparticles (NPs) on Trigonella foenum (fenugreek) in vitro cultures was studied within 3 weeks (on days 1, 7, 14, and 21) and compared with the control and bulk (macrometer-sized particles) alumina-treated groups. Transmission electron microscopy (TEM) and dynamic light scattering were used for the characterization of NPs. The results of TEM analysis represented a nearly spherical shape for the NPs with agglomeration. The zeta potential of alumina NPs was - 25.4 ± 2.5 mV and the averaged diameter was 20 ± 5 nm. Atomic absorption and inductively coupled plasma-optical emission spectroscopy provided evidence for the release and uptake of aluminum. Treatment of cultures with NPs led to an increase in the formation of lateral roots. Treatment of fenugreek with bulk alumina caused a significant decrease in the number of leaves on day 21 (p < 0.05) and the root length on days 14 and 21 compared with the control group (p < 0.05). Alumina NP has led to a significant increase in the malondialdehyde content on days 7, 14, and 21 (p < 0.001). The glutathione content was decreased significantly in NP and bulk-treated groups on days 1 and 7 (p < 0.05). FRAP assay results showed that NPs and bulk alumina led to a decrease in the antioxidant power on days 7, 14, and 21 (p < 0.001). The increased activity of catalase (p < 0.001) and ascorbate peroxidase (p < 0.001) was observed in the bulk-treated group. Lignin content had a significant increase in response to NPs on days 14 and 21 (p < 0.001). Conclusively, alumina nano/macro particles affected agronomical and physiological properties of T. foenum; however, smaller sized particles do not necessarily imply greater toxicity, while uptake of the aluminum ions should be considered seriously.


Using CRP to predict anastomotic leakage after open and laparoscopic colorectal surgery: is there a difference?
C-reactive protein (CRP) has proven to be a useful adjunct in early diagnosis of anastomotic leak (AL) after colorectal surgery. It would be of considerable value to examine whether modality of surgery has influence upon postoperative CRP serum levels and their predictive value in the diagnosis of AL. All patients undergoing elective colorectal surgery with anastomosis were enrolled into a prospective database between 2011 and 2014. AL was defined with strict operative and radiological criteria. Outcomes between open and laparoscopic resections were assessed statistically and Receiver Operating Characteristic (ROC) curve analysis performed. Seven hundred twenty-seven patients with an intestinal anastomosis were identified including 468 laparoscopic procedures (468/727; 64 %). There were 58 anastomotic leaks (58/727; 7.9 %) of which 29 (6.2 %) were laparoscopic and 29 (11.2 %) were open. Mean CRP levels were significantly higher in patients after open surgery compared with laparoscopic both with AL (p = 0.013), and without (p = 0.02). ROC curve analysis revealed postoperative day 3 (cut-off CRP 209) and day 4 (cut-off CRP 123.5) to be most predictive of leak in the open group with an area under the curve (AUC) 0.794 (sensitivity 80 %, specificity 80 %) and AUC 0.806 (sensitivity 94 %, specificity 60 %), respectively. In the laparoscopic group, day 2 proved to be the most accurate day for detection of leak with a cut-off CRP of 146.5 showing 75 % sensitivity and a 70 % specificity (AUC 0.766). CRP levels are higher after open surgery compared with laparoscopic surgery, both with and without AL. AL generates a significant detectable increase in CRP within 2-4 days after surgery.


[Studies of the regulation potential of glucose metabolism in the calf following infusion of fructose solution and using various hormones (ACTH, prednisolone, adrenaline and noradrenaline)].
Infusion of fructose solution (25 g in 100 ml) in sugar rations of 0.5 g or 1.0 g per kilogram of body weight usually induced in calf temporary drop of glucose in the blood plasma. This was attributable to rise in insulin secretion and glucose conversion or insufficiently fast effectiveness of insulin antagonists (adrenaline, glucagon, glucocorticosteroids). Such glucose drop could not be prevented by administration of 50 mg of prednisolone four hours before fructose infusion and 25 IU of ACTH five hours before infusion. Glucose as well as free fatty acids in the blood plasma went up in response to injection of 2.5 microgram/kg body weight during hypoglycaemia. The action of adrenaline was found to depend on the insulin dose given before in cases of insulin-induced hypoglycaemia. Doses of 0.5 g fructose per kilogram of body weight were administered in nine experimental infusions. Average half-life recorded from the blood plasma was 21.3 +/- 3.4 minutes.


Relationship between self-defeating humor and the Gray matter volume in the orbital frontal cortex: the moderating effect of divergent thinking.
The self-defeating humor style is characterized by the excessive use of self-mockery, fawning over others and ineffective denial of negative emotions. The differences in brain structures responsible for self-defeating humor and the role of divergent thinking (DT) in this relationship have not been directly investigated in a large sample. Using voxel-based morphometry (VBM), we identified the association between regional gray matter volume (GMV) and self-defeating humor (assessed by the Humor Style Questionnaire) in 284 participants. Then, the role of DT (assessed by the Torrance Tests of Creative Thinking, TTCT) in the relationship between humor and the related brain regions was examined in the participants (N = 280). The results showed that self-defeating humor was significantly positively associated with the regional GMV in the left orbital frontal cortex (OFC) and that DT had a moderating effect on this relationship. Among the individuals with higher DT, a strong positive correlation was observed between self-defeating humor and the OFC, but among individuals with lower DT, this correlation was weaker. These results reveal that the interaction between brain structures and DT plays an important role in humor, thus providing new evidence enhancing our understanding of the mechanism of humor.


Research of exosome in bone metastasis through dual aptamer recognition based entropy-driven amplification.
Sensitive and accurate detection of exosome will greatly facilitate the early diagnosis of diverse diseases, such as cancers. Herein, a novel dual aptamer recognition based entropy-driven amplification was established for accurate analysis of exosomes. There are two main procedures in the proposed biosensor, including dual aptamer based recognition of exosome and entropy-driven catalytic system based signal recycling. In the recognition process, designed SMBs-S1 probe and S2-S4 probe complex, containing a CD63 aptamer and an EpCAM aptamer, respectively, are utilized for cooperated identification of exosomes. S4 probe was then released from S2-S4 probe complex through chain replacement of S5. The released S4 probe triggers entropy-driven catalytic system based signal recycling and endow the method a superior sensitivity. Impressively, owing to the cooperated identification of CD63 and EpCAM protein, the method exhibited a superior specificity and stayed stable under the interference of free CD63 and/or EpCAM protein. We believe that the sensitive, accurate strategy will provide a powerful tool for multiple biomarkers analysis and related clinical applications.


Clinical Adoption of Advanced Therapies: Challenges and Opportunities.
As the cell and gene therapy field matures the powerful therapeutic potential of these innovative therapies is starting to be shown, particularly in the fields of oncology and childhood immune deficiency diseases. However, as more therapies enter late stage clinical trials, advances and innovation are required in manufacturing, logistics, regulation, reimbursement and the healthcare setting to ensure that systems are in place to support wider clinical adoption of these promising treatments. A window of opportunity exists to implement new methodologies for best practice in both the ability to manufacture products reproducibly at scale, as well as ensuring healthcare systems are not overwhelmed by the variety and complexity of these new therapies and the additional burden they will place on already stretched facilities. If all interested parties work together it will be possible for the sector to develop the necessary processes, skilled staff and infrastructure needed as more treatments move from clinical trial to marketed products.


Activation of peroxisome proliferator-activated receptor gamma bypasses the function of the retinoblastoma protein in adipocyte differentiation.
The retinoblastoma protein (pRB) is an important regulator of development, proliferation, and cellular differentiation. pRB was recently shown to play a pivotal role in adipocyte differentiation, to interact physically with adipogenic CCAAT/enhancer-binding proteins (C/EBPs), and to positively regulate transactivation by C/EBPbeta. We show that PPARgamma-mediated transactivation is pRB-independent, and that ligand-induced transactivation by PPARgamma1 present in RB+/+ and RB-/- mouse embryo fibroblasts is sufficient to bypass the differentiation block imposed by the absence of pRB. The differentiated RB-/- cells accumulate lipid and express adipocyte markers, including C/EBPalpha and PPARgamma2. Interestingly, adipose conversion of pRB-deficient cells occurs in the absence of compensatory up-regulations of the other pRB family members p107 and p130. RB+/+ as well as RB-/- cells efficiently exit from the cell cycle after completion of clonal expansion following stimulation with adipogenic inducers. We conclude that ligand-induced activation of endogenous PPARgamma1 in mouse embryo fibroblasts is sufficient to initiate a transcriptional cascade resulting in induction of PPARgamma2 and C/EBPalpha expression, withdrawal from the cell cycle, and terminal differentiation in the absence of a functional pRB.


Microbial contamination of contact lens care systems.
We examined the contact lens care systems of 100 asymptomatic patients who used hard or soft contact lenses for correction of refractive errors for the presence of bacteria, fungi, Acanthamoeba, and endotoxin. Of 100 patients, 52 had contaminated contact lens care systems. Sixteen of 126 bottles (13%) of commercial contact lens care solutions were contaminated. Contaminated commercial solutions were opened and used for a longer period of time than uncontaminated solutions. Contamination was not found in bottles of preserved commercial solutions that were opened and used for less than 21 days. All 12 bottles of homemade saline were contaminated with bacteria, and Acanthamoeba was isolated from two of these bottles. Pseudomonas was found in the care systems of 12 patients. Bacillus species, which form spores resistant to heat, were found in the care systems of seven patients. Endotoxin, which is also resistant to heat, was detected in nine of 35 care systems (26%) tested. Potential pathogens were isolated from the conjunctiva of six patients.


A method for correctly setting the rf flip angle.
Currently the accepted method for setting the correct rf power levels to achieve 90 degrees and 180 degrees rf pulses for MR imaging is to peak the echo amplitude of a rf spin-echo sequence. The echo amplitude of this alpha-2 alpha pulse sequence is proportional to sin3 (alpha) and has a relatively broad maximum. Recently another method for setting the rf flip angle by maximizing the ratio of the stimulated echo to the primary echo amplitudes (in a 3 alpha sequence) demonstrated accuracy similar to that of the spin-echo method using a shorter repetition time. We present a new, more sensitive, and more accurate method for setting the correct rf power levels for 90 degrees and 180 degrees rf pulses. In this method, based upon the stimulated echo pulse sequence, we are able to accurately set the rf power to within +/- 0.1 dB by minimizing the signal amplitude of the third spin echo. This null method works for both selective and nonselective rf pulses of flip angle 90 degrees or 180 degrees, allowing the user to accurately adjust the relative amplitudes of the four rf pulse types within a single pulse sequence.


Particle radiotherapy in the era of radioimmunotherapy.
Radiotherapy (RT) is one of the key modalities for cancer treatment, and more than 70% of tumor patients will receive RT during the course of their disease. Particle radiotherapy, such as proton radiotherapy, carbon-ion radiotherapy (CIRT) and boron neutron capture therapy (BNCT), is currently available for the treatment of patients Immunotherapy combined with photon RT has been successfully used in the clinic. The effect of immunotherapy combined with particle RT is an area of interest. However, the molecular mechanisms underlying the effects of combined immunotherapy and particle RT remain largely unknown. In this review, we summarize the properties of different types of particle RT and the mechanisms underlying their radiobiological effects. Additionally, we compared the main molecular players in photon RT and particle RT and the mechanisms involved the RT-mediated immune response.


Cocaine produces fine structural nuclear alterations in cultured neuroglioblastoma cells.
NG 108-15 neuroglioblastoma cells were grown in culture medium containing either 10(-3), 10(-6) or 10(-9) M cocaine for 1-3 days. Some cultures were assessed for viability while others were processed for electron microscopy. Following 1-3 days of cocaine, no cytoplasmic alterations were observed compared to control; however, numerous dense bodies were present in some cells cultured with the highest doses. After 2 days of treatment with 10(-3) and 10(-6) M cocaine, nuclear invaginations were found filled with vesicles, and the nuclear membrane surrounding the vesicles was disrupted. Following 3 days of treatment with 10(-3) and 10(-6) M cocaine, patches of vesicles and tubules were also seen in the nucleus, but without surrounding membranes. The vesicles ranged in size from 0.05 to 0.8 micron. Cell counts revealed a significant slowing in the rate of cell division after two days of exposure to 10(-3) M cocaine. All concentrations of cocaine caused a significant decrease in cell viability by the third day of treatment. These results suggest that cocaine may interfere with cell replication and also may have a neurotoxic effect.


Functionally distinct human T cell clones that produce lymphokines with IL-2-like activity.
Various functionally distinct human T cell clones derived from in vitro mixed leukocyte cultures are found to secrete lymphokines with detectable Interleukin-2 (IL-2)-like activity upon antigenic stimulation. These lymphokine producing clones are not only dependent on exogenous growth factors provided in the form of crude phytohemagglutinin (PHA)-induced spleen conditioned medium for growth but can themselves be driven to proliferate by their own lymphokines. The induction of lymphokine production appears to be antigen-specific and the lymphokines secreted are believed to contain nonantigen and nonspecies specific IL-2-like activity. We show here that IL-2-like lymphokines are produced by a subset of T lymphocyte clones, i.e., the help-independent cytotoxic T cells clones that have the capacity to proliferate to, as well as to lyse, the original sensitizing cells. In addition, other T cell clones capable of producing active lymphocytes include those clones that have the T helper cell characteristics, i.e., can undergo antigen-induced proliferation but are not cytotoxic; in the presence of lectin (PHA), however, some helper T cells (Th) but not others, can express lectin-dependent cell-mediated lysis. Finally, yet another subset of T lymphocytes, the help-dependent cytotoxic T cell clones that cannot proliferate to antigenic stimulation, was found to produce no detectable IL-2-like activity.


The vasculature niches required for hematopoiesis.
Endothelial cells play a critical role in supporting postnatal hematopoiesis in the bone marrow. Unique endothelial cells, together with various perivascular cells, form different types of vascular structures, constructing a vast microvascular delivery and trafficking network for blood cells, oxygen, and nutrition. These blood vessels build distinct hematopoietic stem and progenitor cell niches, which offer not only sites of residence for blood cells but also indispensable signals directing HSPC homing, self-renewal, and multilineage differentiation. Deep insight into the structure and function of the BM vasculature niche and its participation in hematopoiesis is necessary to develop advanced strategies for the reconstitution of hematopoiesis.


Romantic functioning and depressive symptoms among early adolescent girls: the moderating role of parental emotional availability.
This study tested associations between adolescent romantic functioning and depressive symptoms and predicted that adolescents with emotionally unavailable parents would be most likely to show an association between poor romantic functioning and depressive symptoms. Data collected from 80 early adolescent nonreferred girls (average age of 13.45; SD = .68) and their mothers supported predictions. A higher frequency of romantic experiences (e.g., going on dates) and poor romantic competence were each associated with higher levels of current depressive symptoms. Moreover, these associations were most strong for girls with emotionally unavailable parents.


Counterbalancing effects of maternal mercury exposure during different stages of early ontogeny in American toads.
Maternal transfer of environmental contaminants is a disadvantageous parental effect which can have long-lasting implications for offspring fitness. We investigated the effects of mercury (Hg) on the reproductive success of female amphibians and the subsequent effects of maternal transfer on the development of their offspring. American toads (Bufo americanus) maternally transferred Hg to their eggs, and there was a negative relationship between Hg concentrations and the percentage of viable hatchlings produced in clutches. However, when we continued to monitor larvae that successfully hatched, we found 21% greater metamorphic success in larvae from Hg-exposed mothers compared to reference larvae. The negative effect in the embryonic stage and positive effect in the larval stage counterbalanced one another, ultimately resulting in no difference in predicted terrestrial recruitment, regardless of maternal Hg exposure. Our findings demonstrate that maternal effects on survival manifesting at different stages in ontogeny have the potential to produce complicated outcomes.


Hormone-sensitive lipase is involved in hepatic cholesteryl ester hydrolysis.
Hormone-sensitive lipase (HSL) regulates the hydrolysis of acylglycerol and cholesteryl ester (CE) in various organs, including adipose tissues. However, the hepatic expression level of HSL has been reported to be almost negligible. In the present study, we found that mice lacking both leptin and HSL (Lep(ob/ob)/HSL(-/-)) showed massive accumulation of CE in the liver compared with Lep(ob/ob)/HSL(+/+) mice, while triacylglycerol (TG) accumulation was modest. Similarly, feeding with a high-cholesterol diet induced hepatic CE accumulation in HSL(-/-) mice. Supporting these observations, we detected significant expression of protein as well as mRNA of HSL in the liver. HSL(-/-) mice showed reduced activity of CE hydrolase, but not of TG lipase, in the liver compared with wild-type mice. Furthermore, we confirmed the expression of HSL in viable parenchymal cells isolated from wild-type mice. The hepatocytes from HSL(-/-) mice showed reduced activity of CE hydrolase and contained more CE than those from HSL(+/+) mice even without the incubation with lipoproteins. Incubation with LDL further augmented the accumulation of CE in the HSL-deficient hepatocytes. From these results, we conclude that HSL is involved in the hydrolysis of CE in hepatocyes.


Regensburg Insomnia Scale (RIS): a new short rating scale for the assessment of psychological symptoms and sleep in insomnia; study design: development and validation of a new short self-rating scale in a sample of 218 patients suffering from insomnia and 94 healthy controls.
The Regensburg Insomnia Scale (RIS) is a new self-rating scale to assess cognitive, emotional and behavioural aspects of psychophysiological insomnia (PI) with only ten items. A specific purpose of the new scale is the evaluation of the outcome of insomnia- specific cognitive behaviour therapy (CBT-I). Internal consistency of the RIS has been validated in 218 patients with PI. For determining sensitivity and specificity, this sample has been compared to 94 healthy controls. Sensitivity to change and pre-post cross-validation with the Pittsburgh Sleep Quality Index (PSQI) has been tested in a separate sample of 38 patients with PI undergoing CBT-I. RIS distinguishes well between controls and patients with PI. Internal consistency was within a good range (Cronbach alpha = .890). RIS was sensitive for detecting improvements after CBT-I in sleep parameters and target symptoms such as sleep-related thinking. The RIS is a valid and feasible instrument for assessing psychological PI-symptoms and sleep parameters.


Pediatric germ cell tumors.
Pediatric germ cell tumors are a diverse group of neoplasms with variable clinical behaviors, depending upon the age and site of presentation. Most result from sporadic mutations, although environmental exposures and other genetic aberrations may play a role. Platinum-based chemotherapy has dramatically improved the event-free and overall survival outcomes of pediatric patients with malignant germ cell tumors over the past two decades. Prognosis is dependent on tumor stage and location. Patients with gonadal germ cell tumors have at least a 95% 5-year survival for early stage disease and at least a 85% 5-year survival for advanced stages. In general, extragonadal germ cell tumors carry a poorer prognosis with mediastinal location having the worst outcomes (70% 4-year survival). Current trials are focused on maintaining similar excellent outcomes while reducing morbidity by reducing the dose and duration of chemotherapy. Cytogenetic research studies have found chromosomal aberrations specific to some of these tumors that may serve as prognosticators and even direct therapy.


Rheumatoid arthritis--a sequel to HBsAg hepatitis.
Reported here is the first case of classic rheumatoid arthritis emerging in the setting of hepatitis B surface antigen (HBsAG)-positive viral hepatitis. Polyfocal arthritis and myalgia, lymphadenopathy and constitutional symptoms were the presenting manifestations of anicteric hepatitis in this 23 year old man. Smooth muscles antibodies, HBsAg and "rheumatoid" factor were demonstrated initially. The hepatocellular disease, biopsy-proved, resolved completely and without recurrence; clinically and serologically. Latex test positivity persisted, increasing in titer, and polyarthritis progressed to fulfull criteria for classic rheumatoid arthritis, with erosions on roentgenogram and characteristic synovial disease. After 60 months of follow-up, the rheumatoid synovitis has continued to progress despite appropriate therapy. The arthritis of viral hepatitis and the significance of rheumatoid factor in association with hepatocellular disease are discussed.


XitoSBML: A Modeling Tool for Creating Spatial Systems Biology Markup Language Models From Microscopic Images.
XitoSBML is a software tool designed to create an SBML (Systems Biology Markup Language) Level 3 Version 1 document from microscopic cellular images. It is implemented as an ImageJ plug-in and is designed to create spatial models that reflect the three-dimensional cellular geometry. With XitoSBML, users can perform spatial model simulations based on realistic cellular geometry by using SBML-supported software tools, including simulators such as Virtual Cell and Spatial Simulator. XitoSBML is open-source and is available at https://github.com/spatialsimulator/XitoSBML/. XitoSBML is confirmed to run on most 32/64-bit operating systems: Windows, MacOS, and Linux.


Evidence for a mechanism predisposing to intergenerational CAG repeat instability in spinocerebellar ataxia type I.
Spinocerebellar ataxia type I (SCAI) is an autosomal dominant neurodegenerative disease caused by the expansion of a CAG trinucleotide repeat on chromosome 6p. Normal alleles range from 19-36 repeats while SCA1 alleles contain 43-81 repeats. We now show that in 63% of paternal transmissions, an increase in repeat number is observed, whereas 69% of maternal transmissions showed no change or a decrease in repeat number. Sequence analysis of the repeat from 126 chromosomes reveals an interrupted repeat configuration in 98% of the unexpanded alleles but a contiguous repeat (CAG)n configuration in 30 expanded alleles from seven SCA1 families. This indicates that the repeat instability in SCA1 is more complex than a simple variation in repeat number and that the loss of an interruption predisposes the SCA1 (CAG)n to expansion.


Experimental study on tumor affinity of 201T1-chloride.
The tumor affinity of 201T1 was studied with normal and VX-2 cancer-bearing rabbits. 201T1 distribution in normal rabbit tissues was greatest in the kidney and heart muscle, followed by thyroid gland small intestine, spleen, lung, liver, bone marrow, bone, skeletal muscle, and blood, respectively. Accumulation into the thyroid varied greatly according to individuals. Generally, the taller the height of follicular cells, the greater was the affinity. Accumulation of 201T1 into the tumor transplanted into femoral muscle reached its maximum within 1 h after administration, and thereafter decreased gradually. When the tumor affinity was compared with that of 67Ga, the ratio of 67Ga accumulation to tissues (except blood) was greater than that of 201T1. Accumulation of 201T1 is significantly correlated to that of 42K, and the mechanism of 201T1-tumor affinity seems to be triggered by the acceleration of the potassium metabolism of a tumor. Accumulation into the inflammatory focus was greater with 67Ga as a ratio to muscle, while the ratio to blood was greater with 201T1.


Ultrahigh-field magnetic resonance imaging: the clinical potential for anatomy, pathogenesis, diagnosis, and treatment planning in brain disease.
In this review, current (clinical) applications and possible future directions of ultrahigh-field (≥7 T) magnetic resonance (MR) imaging in the brain are discussed. Ultrahigh-field MR imaging can provide contrast-rich images of diverse pathologies and can be used for early diagnosis and treatment monitoring of brain disease. These images may provide increased sensitivity and specificity. Several limitations need to be overcome before worldwide clinical implementation can be commenced. Current literature regarding clinically based ultrahigh-field MR imaging is reviewed, and limitations and promises of this technique are discussed, as well as some practical considerations for the implementation in clinical practice.


Incorporation of natriuretic peptides with clinical risk scores to predict heart failure among individuals with dysglycaemia.
To evaluate the performance of the WATCH-DM risk score, a clinical risk score for heart failure (HF), in patients with dysglycaemia and in combination with natriuretic peptides (NPs). Adults with diabetes/pre-diabetes free of HF at baseline from four cohort studies (ARIC, CHS, FHS, and MESA) were included. The machine learning- [WATCH-DM(ml)] and integer-based [WATCH-DM(i)] scores were used to estimate the 5-year risk of incident HF. Discrimination was assessed by Harrell's concordance index (C-index) and calibration by the Greenwood-Nam-D'Agostino (GND) statistic. Improvement in model performance with the addition of NP levels was assessed by C-index and continuous net reclassification improvement (NRI). Of the 8938 participants included, 3554 (39.8%) had diabetes and 432 (4.8%) developed HF within 5 years. The WATCH-DM(ml) and WATCH-DM(i) scores demonstrated high discrimination for predicting HF risk among individuals with dysglycaemia (C-indices = 0.80 and 0.71, respectively), with no evidence of miscalibration (GND P ≥0.10). The C-index of elevated NP levels alone for predicting incident HF among individuals with dysglycaemia was significantly higher among participants with low/intermediate (<13) vs. high (≥13) WATCH-DM(i) scores [0.71 (95% confidence interval 0.68-0.74) vs. 0.64 (95% confidence interval 0.61-0.66)]. When NP levels were combined with the WATCH-DM(i) score, HF risk discrimination improvement and NRI varied across the spectrum of risk with greater improvement observed at low/intermediate risk [WATCH-DM(i) <13] vs. high risk [WATCH-DM(i) ≥13] (C-index = 0.73 vs. 0.71; NRI = 0.45 vs. 0.17). The WATCH-DM risk score can accurately predict incident HF risk in community-based individuals with dysglycaemia. The addition of NP levels is associated with greater improvement in the HF risk prediction performance among individuals with low/intermediate risk than those with high risk.


Inhibiting IDO pathways to treat cancer: lessons from the ECHO-301 trial and beyond.
With immunotherapy enjoying a rapid resurgence based on the achievement of durable remissions in some patients with agents that derepress immune function, commonly referred to as "checkpoint inhibitors," enormous attention developed around the IDO1 enzyme as a metabolic mediator of immune escape in cancer. In particular, outcomes of multiple phase 1/2 trials encouraged the idea that small molecule inhibitors of IDO1 may improve patient responses to anti-PD1 immune checkpoint therapy. However, recent results from ECHO-301, the first large phase 3 trial to evaluate an IDO1-selective enzyme inhibitor (epacadostat) in combination with an anti-PD1 antibody (pembrolizumab) in advanced melanoma, showed no indication that epacadostat provided an increased benefit. Here we discuss several caveats associated with this failed trial. First is the uncertainty as to whether the target was adequately inhibited. In particular, there remains a lack of direct evidence regarding the degree of IDO1 inhibition within the tumor, and previous trial data suggest that sufficient drug exposure may not have been achieved at the dose tested in ECHO-301. Second, while there is a mechanistic rationale for the combination tested, the preclinical data were not particularly compelling. More efficacious combinations have been demonstrated with DNA damaging modalities which may therefore be a more attractive alternative. Third, as a highly selective IDO1 inhibitor, epacadostat was advanced aggressively despite preclinical genetic evidence of tumors bypassing IDO1 blockade. Indeed, a well-grounded literature starting in 2011 points to targeting strategies that account for both IDO and tryptophan 2,3-dioxygenase as more appealing directions to pursue, including dual inhibitors and inhibitors of nodal downstream effector pathways such as aryl hydrocarbon receptor blockade. Overall, the clinical readout from a single trial with significant limitations is by no means a definitive test for the field. While biomarker information yet to be gleaned from ECHO-301 may yet reveal useful information regarding IDO1 pathway drugs, better rationalized compounds and better rationalized trial designs will be important in the future to accurately gauge medical impact.


Microplastic distribution in different tissues of small pelagic fish of the Northeast Atlantic Ocean.
The accumulation of microplastics (MP) by marine species of ecological and commercial interest represents a major concern, particularly for those present in human diet. This study analysed the accumulation of MP in three species of coastal pelagic fish with high commercial value, European sardine (Sardina pilchardus), European anchovy (Engraulis encrasicolus) and horse mackerel (Trachurus trachurus), collected along the Western coast of the Iberian Peninsula. The gastrointestinal tract (GT), gills and muscle were analysed and a total of 504 particles were observed. MP were found in all target tissues of the studied species. Horse mackerel exhibited significantly higher concentrations of microplastics in GT compared to other tissues. On the other hand, anchovies and sardines had significantly lower microplastic concentrations in their muscle tissue. The accumulation of microplastics in the gills showed a significant difference between species, with anchovy having significantly higher concentrations compared to horse mackerel. Horse mackerel had the highest percentage of individuals with microplastics in their GT (92 %), followed by sardine (75 %) and anchovy (50 %). Horse mackerel was also the species that registered the highest percentage of individuals with particles in the muscle (63 %), followed by anchovy (40 %) and finally sardine (39 %). MP in the gills of European sardines and anchovies were similar to those found in water samples. The majority of MP found measured <0.5 mm and were blue fibers. Furthermore, the presence of MP in the GT showed a weak and moderated significant negative correlation with the Fulton Condition Index in horse mackerel and European sardine. Our study confirms the ubiquitous extent of MP contamination in the ocean and provides baseline evidence of MP tissue distribution in three small pelagic fish species with distinct feeding behaviour, while correlating this with the presence of MP in water. Importantly, the results of this study contribute to improve the understanding of biological partitioning of MP in open sea fish species with high commercial relevance, and the potential deleterious effects of our increasingly MP contaminated world.


Towards MR contrast independent synthetic CT generation.
The use of synthetic CT (sCT) in the radiotherapy workflow would reduce costs and scan time while removing the uncertainties around working with both MR and CT modalities. The performance of deep learning (DL) solutions for sCT generation is steadily increasing, however most proposed methods were trained and validated on private datasets of a single contrast from a single scanner. Such solutions might not perform equally well on other datasets, limiting their general usability and therefore value. Additionally, functional evaluations of sCTs such as dosimetric comparisons with CT-based dose calculations better show the impact of the methods, but the evaluations are more labor intensive than pixel-wise metrics. To improve the generalization of an sCT model, we propose to incorporate a pre-trained DL model to pre-process the input MR images by generating artificial proton density, T1 and T2 maps (i.e. contrast-independent quantitative maps), which are then used for sCT generation. Using a dataset of only T2w MR images, the robustness towards input MR contrasts of this approach is compared to a model that was trained using the MR images directly. We evaluate the generated sCTs using pixel-wise metrics and calculating mean radiological depths, as an approximation of the mean delivered dose. On T2w images acquired with the same settings as the training dataset, there was no significant difference between the performance of the models. However, when evaluated on T1w images, and a wide range of other contrasts and scanners from both public and private datasets, our approach outperforms the baseline model. Using a dataset of T2w MR images, our proposed model implements synthetic quantitative maps to generate sCT images, improving the generalization towards other contrasts. Our code and trained models are publicly available.


Reduced central sympathetic activity in Parkinson's disease.
With a combination of different sympathetic tests, we aimed to elucidate whether impairment of sympathetic function in Parkinson's disease (PD) is the consequence of a central or peripheral efferent dysfunction. Thirty-five patients with early-to-intermediate PD (median age: 63 years; IQR: 57-67 years; disease duration 1-9 years, 15 women) and 20 age- and sex-matched healthy controls (median age: 64.5 years; IQR: 58-68 years; 10 women) were recruited. Autonomic testing was performed in two subgroups and included the assessment of resting cardiovascular parameters, postprandial hypotension (PPH), orthostatic hypotension (OH), and vasoconstriction induced by intradermal microdialysis with different concentrations of norepinephrine (NE; 10-5 ; 10-6 ; 10-7 ; 10-8 ) and by cold through forehead cooling. We also used sympathetic multiunit microneurography (muscle sympathetic nerve activity; MSNA; burst frequency (BF): bursts per minute; burst incidence (BI): bursts per 100 heart beats) and evaluated the presence of phosphorylated α-synuclein deposits in skin innervation in biopsies from the thighs by immunohistohemistry. Diastolic blood pressure was higher in the PD group at rest (p < .001) and during OH (F = 6.533; p = .022). Vasoconstriction induced by NE microdialysis and cold was unchanged in PD patients. MSNA was lower in PD patients than in controls (BF: p = .001; BI: p = .025). Phosphorylated α-synuclein deposits could be found only in PD patients. We did not find indications for peripheral sympathetic nerve fiber dysfunction or adrenoreceptor sensitivity changes. The decreased MSNA argues in favor of central sympathetic impairment.


Comparison of the Pentacam equivalent keratometry reading and IOL Master keratometry measurement in intraocular lens power calculations.
To compare the accuracy of the Pentacam Holladay equivalent keratometry readings with the IOL Master 500 keratometry in calculating intraocular lens power. Non-randomized, prospective clinical study conducted in private practice. Forty-five consecutive normal patients undergoing cataract surgery. Forty-five consecutive patients had Pentacam equivalent keratometry readings at the 2-, 3 and 4.5-mm corneal zone and IOL Master keratometry measurements prior to cataract surgery. For each Pentacam equivalent keratometry reading zone and IOL Master measurement the difference between the observed and expected refractive error was calculated using the Holladay 2 and Sanders, Retzlaff and Kraff theoretic (SRKT) formulas. Mean keratometric value and mean absolute refractive error. There was a statistically significantly difference between the mean keratometric values of the IOL Master, Pentacam equivalent keratometry reading 2-, 3- and 4.5-mm measurements (P < 0.0001, analysis of variance). There was no statistically significant difference between the mean absolute refraction error for the IOL Master and equivalent keratometry readings 2 mm, 3 mm and 4.5 mm zones for either the Holladay 2 formula (P = 0.14) or SRKT formula (P = 0.47). The lowest mean absolute refraction error for Holladay 2 equivalent keratometry reading was the 4.5 mm zone (mean 0.25 D ± 0.17 D). The lowest mean absolute refraction error for SRKT equivalent keratometry reading was the 4.5 mm zone (mean 0.25 D ± 0.19 D). Comparing the absolute refraction error of IOL Master and Pentacam equivalent keratometry reading, best agreement was with Holladay 2 and equivalent keratometry reading 4.5 mm, with mean of the difference of 0.02 D and 95% limits of agreement of -0.35 and 0.39 D. The IOL Master keratometry and Pentacam equivalent keratometry reading were not equivalent when used only for corneal power measurements. However, the keratometry measurements of the IOL Master and Pentacam equivalent keratometry reading 4.5 mm may be similarly effective when used in intraocular lens power calculation formulas, following constant optimization.


Application of molecular dynamics simulation in self-assembled cancer nanomedicine.
Self-assembled nanomedicine holds great potential in cancer theragnostic. The structures and dynamics of nanomedicine can be affected by a variety of non-covalent interactions, so it is essential to ensure the self-assembly process at atomic level. Molecular dynamics (MD) simulation is a key technology to link microcosm and macroscale. Along with the rapid development of computational power and simulation methods, scientists could simulate the specific process of intermolecular interactions. Thus, some experimental observations could be explained at microscopic level and the nanomedicine synthesis process would have traces to follow. This review not only outlines the concept, basic principle, and the parameter setting of MD simulation, but also highlights the recent progress in MD simulation for self-assembled cancer nanomedicine. In addition, the physicochemical parameters of self-assembly structure and interaction between various assembled molecules under MD simulation are also discussed. Therefore, this review will help advanced and novice researchers to quickly zoom in on fundamental information and gather some thought-provoking ideas to advance this subfield of self-assembled cancer nanomedicine.


The metabolism of ribosomal proteins microinjected into the oocytes of Xenopus laevis.
When the total proteins from Xenopus laevis 60 S ribosomal subunits (TP60) were 3H-labeled in vitro and injected back into X. laevis oocytes, most 3H-TP60 are integrated into the cytoplasmic 60 S subunits via the nucleus during 16 h of incubation. In the oocytes whose rRNA synthesis is inhibited, 3H-TP60 are rapidly degraded with a half-life of 2-3 h. This degradation ceased as soon as rRNA synthesis was resumed, suggesting that ribosomal proteins unassociated with nascent rRNA are unstable in the oocytes. The degradation of 3H-TP60 in the absence of RNA synthesis was inhibited by iodoacetamide, a cysteine protease inhibitor, resulting in the accumulation of 3H-TP60 in the nucleus reaching about a threefold concentration in the cytoplasm. Considering the results with enucleated oocytes, we suggest that the X. laevis nucleus has a limited capacity to accumulate ribosomal proteins in an active manner but that those ribosomal proteins accumulated in excess over rRNA synthesis are degraded by a cysteine protease in the nucleus. By contrast, ribosomal proteins from Escherichia coli only equilibrate between the nucleus and the cytoplasm and are degraded by serine protease(s) in the cytoplasm without being integrated in the form of ribosomes in the nucleus.


On Navigating Analytical Choices in Research on Early Life Adversity: A Commentary on Sisitsky et al. (2023).
Early life adversities (ELA), including exposure to childhood maltreatment, deprivation or community violence, rarely occur in isolation. This co-occurrence poses several conceptual and methodological challenges for researchers, who must decide how best to model ELA and its association with outcomes. In this commentary, we discuss how different analytical choices come with their own - often complementary - sets of assumptions, strengths and limitations, which should be carefully considered when designing research on ELA. We then summarize work published in this issue by Sisitsky et al. (Research on Child and Adolescent Psychopathology, 2023), which serves as an important example of how different approaches can be incorporated in research in order to capture ELA as a complex phenomenon, while generating actionable results. Ultimately, such integration can enhance the quality and relevance of research, contributing to a more comprehensive understanding of ELA and its effects on health outcomes, paving the way for more targeted prevention and intervention strategies to promote children's wellbeing.


Novel imaging techniques reshape the landscape in high-risk prostate cancers.
High-risk prostate cancers (PCa), that is, those with prostate-specific antigen greater than 20 ng/dl, Gleason Score of at least 8, or extraprostatic spread, are nowadays commonly treated by surgery and radiotherapy combined with a fixed period of systemic treatment. Implementing these strategies requires an exhaustive assessment of metastatic spread. This review addresses the latest development in integrated imaging techniques. In contrast to the progress that has been made in PCa treatment, diagnostic strategies have not much evolved. Most guidelines still recognize (99m)Tc bone scintigraphy and computed tomography (CT) as cornerstone modalities to assess metastatic spread in bones and lymph nodes. Therefore, modern imaging techniques should primarily focus on these two targets. PET with various tracers, including (11)C or (18)F-choline and (18)F-sodium fluoride, and MRI with or without diffusion-weighted imaging are competing to supplant bone scan and CT scan as reference imaging techniques. This review focuses on the latest development of these techniques and analyses their potential impact in everyday urology practice. Although certain hurdles remain, PET and whole-body MRI have the ability to supplant (99m)Tc bone scan and CT as upfront test to assess metastatic spread in high-risk PCa.


Efficacy and tolerance of acne treatment using both spironolactone and a combined contraceptive containing drospirenone.
The use of either oral contraceptives or antiandrogens has been shown to be helpful in the treatment of women with acne. We investigated the safety and efficacy of acne treatment using both spironolactone and a combined contraceptive containing drospirenone. Twenty-seven women with either severe papular or nodulocystic facial acne were treated with a combined oral contraceptive containing 30 mug ethinyl estradiol and 3 mg drospirenone (EE/DRSP; Yasmin) and spironolactone (SL) 100 mg taken daily. A serum potassium level was obtained before initiation of therapy. Between 4 and 6 weeks after the start of both medications, a second serum potassium level was obtained. Side effects were recorded. No significant elevation of serum potassium was found in any of the subjects nor were there any reported additional side effects significant enough to discontinue treatment. At follow up, 85% of subjects were entirely clear of acne lesions or had excellent improvement, 7.4% were mildly improved, and 7.4% were not improved. This was a small prospective study. The combination of EE/DRSP and SL 100 mg daily appears to have efficacy and is well tolerated in the treatment of severe papular and nodulocystic acne in women.


Cholesterol ester hydrolase(s) in mammalian brain: is there a myelin-specific cholesterol ester hydrolase?
The present study compared the properties of cholesterol ester hydrolase(s) in myelin and microsomes from rat, mouse and human brain. The results indicated that the enzyme activity in both myelin and microsomes from rat, mouse and human brain was optimal at pH 6.5 and required Triton X-100 for optimal activity. The enzyme activity in myelin was 3- to 4-fold higher in the presence of Triton X-100 than taurocholate. Addition of phosphatidyl serine enhanced (2 to 4 fold) the hydrolase activity in both myelin and microsomes. The properties of the enzyme in solubilized preparation of myelin were also similar to the properties of the enzyme in partially delipidated and solubilized preparations of microsomes. The activity was again optimal at pH 6.5, required Triton X-100 for optimal activity and was stimulated by phosphatidyl serine. These results indicate that the properties of cholesterol ester hydrolase in myelin are similar to those of the microsomal enzyme and that this is true for the fractions from both human and rodent brain. The data thus lead us to believe that the hydrolase activity in mammalian brain myelin and microsomes may reflect the distribution of a single enzyme in the two fractions rather than two distinct enzymes, one being specific to each fraction.


Investigation of the Mechanism of Electron Capture and Electron Transfer Dissociation of Peptides with a Covalently Attached Free Radical Hydrogen Atom Scavenger.
The mechanisms of electron capture and electron transfer dissociation (ECD and ETD) are investigated by covalently attaching a free-radical hydrogen atom scavenger to a peptide. The 2,2,6,6-tetramethylpiperidin-l-oxyl (TEMPO) radical was chosen as the scavenger due to its high hydrogen atom affinity (ca. 280 kJ/mol) and low electron affinity (ca. 0.45 ev), and was derivatized to the model peptide, FQXTEMPOEEQQQTEDELQDK. The XTEMPO residue represents a cysteinyl residue derivatized with an acetamido-TEMPO group. The acetamide group without TEMPO was also examined as a control. The gas phase proton affinity (882 kJ/mol) of TEMPO is similar to backbone amide carbonyls (889 kJ/mol), minimizing perturbation to internal solvation and sites of protonation of the derivatized peptides. Collision induced dissociation (CID) of the TEMPO tagged peptide dication generated stable odd-electron b and y type ions without indication of any TEMPO radical induced fragmentation initiated by hydrogen abstraction. The type and abundance of fragment ions observed in the CID spectra of the TEMPO and acetamide tagged peptides are very similar. However, ECD of the TEMPO labeled peptide dication yielded no backbone cleavage. We propose that a labile hydrogen atom in the charge reduced radical ions is scavenged by the TEMPO radical moiety, resulting in inhibition of N-Cα backbone cleavage processes. Supplemental activation after electron attachment (ETcaD) and CID of the charge-reduced precursor ion generated by electron transfer of the TEMPO tagged peptide dication produced a series of b + H (bH) and y + H (yH) ions along with some c ions having suppressed intensities, consistent with stable O-H bond formation at the TEMPO group. In summary, the results indicate that ECD and ETD backbone cleavage processes are inhibited by scavenging of a labile hydrogen atom by the localized TEMPO radical moiety. This observation supports the conjecture that ECD and ETD processes involve long-lived intermediates formed by electron capture/transfer in which a labile hydrogen atom is present and plays a key role with low energy processes leading to c and z ion formation. Ab initio and density functional calculations are performed to support our conclusion, which depends most importantly on the proton affinity, electron affinity and hydrogen atom affinity of the TEMPO moiety.


Postmortem stability of some markers of intra-vital wounds.
We have studied the diagnostic value of several markers of the intra-vital nature of wounds - cathepsin D (EC 22.214.171.124) and ions (Ca, Mg, Cu, Zn and Fe) - after the influence of putrefaction. For this purpose, we have inflicted vital wounds to six pigs, which were killed 20 min later. Ten minutes after death, wounds were excised with 5-6 cm of skin around the incision and maintained at three different temperatures (4, 18 and 28.5 +/- 13.4 degrees C). After varying periods of postmortem interval from 0 to 48 h, aliquots of each wound were taken and analyzed with atomic absorption spectrophotometry for ions and with UV-spectrophotometry for cathepsin D. Our results demonstrate that ions conserve their diagnostic ability to differentiate vital from postmortem wounds after the influence of putrefaction. Nevertheless, cathepsin D does not show this ability in these experimental conditions.


Phase II evaluation of diglycoaldehyde, VP-16-213, and the combination of methyl-CCNU and beta-2'-deoxythioguanosine in previously treated patients with colorectal cancer: an Eastern Cooperative Oncology Group study (EST-1275).
The Eastern Cooperative Oncology Group assessed the activity of diglycoaldehyde (DGA), VP-16-213, and the combination of methyl-CCNU and beta-2'-deoxythioguanosine in previously treated patients with advanced colorectal cancer. Objective responses were seen in two of 40 evaluable patients receiving methyl-CCNU and beta-2'-deoxythioguanosine and in one of 35 patients receiving DGA. None of 33 patients responded to VP-16-213, but one death related to sepsis and bone marrow failure occurred. Survival of patients whose previous chemotherapy included a nitrosourea was markedly shortened compared to those who had not been exposed to nitrosoureas. With the possible exception of DGA, further treatment of patients with colorectal cancer with these therapies is not warranted.


Local catheter-based delivery of antithrombotic or antiproliferative drugs: a new concept for prevention of restenosis.
Drug eluting stents have reduced the incidence of restenosis after percutaneous coronary interventions significantly, but cause concern about long term safety. Local drug delivery using special application catheters is an alternative approach for intracoronary pharmacotherapy. Besides the fact, that no problematic coating as drug carrier has to be used, a local delivery independent of the stent itself by using catheter techniques offers further advantages - such as the possibility to treat the whole vessel wall, stent edges and adjacent vessel segments and not only the area close to the stent struts. We have developed a new local catheter-based delivery system for local intracoronary pharmacotherapy. An antithrombotic as well as an antiproliferative therapy concept for prevention of restenosis are presented in the manuscript. Our data show that local drug delivery of platelet glycoprotein VI and paclitaxel were effective in the reduction of thrombus formation and neointima formation in experimental animal models. A combination of early antithrombotic and antiatherosclerotic mechanisms may be a realistic and effective approach to minimize postinterventional thromboischemic events and neointima formation. These results may contribute to an advanced and even combined local intracoronary pharmacotherapy in near future, independent of stent coatings.


Evaluation of cis-diamminedichloroplatinum (II) (cisplatin) neurotoxicity in rats.
Cisplatin neurotoxicity was studied in rats by using electrophysiological and functional tests. The drug was injected intraperitoneally two times a week (1 mg/kg) up to a cumulative dose of 19.0 mg/kg body wt. At this dose the H-reflex-related sensory nerve conduction velocity was reduced. After cessation of drug administration, the H-reflex-related sensory nerve conduction velocity slowly improved. The motor nerve conduction velocity remained unaffected throughout the intoxication period and concentric needle investigation showed no signs of denervation. In addition, the sensorimotor response to a noxious stimulus presented locally to the foot sole was not affected by the cisplatin treatment. It is concluded that measurement of H-reflex-related sensory nerve conduction velocity is a useful and accurate tool in the evaluation of cisplatin neurotoxicity in rats.


Normalized upper cervical spinal cord atrophy in multiple sclerosis.
To find an optimal normalizing factor for upper cervical spinal cord area (UCCA) and to establish whether, in a cross-sectional study, the normalized UCCA correlates better with the neurological disability than the absolute measurement in multiple sclerosis patients. UCCA and three potential normalizing factors were estimated from magnetic resonance imaging data in 51 control subjects. Their reliability was assessed and the linear relationships between UCCA and three potential correction factors were investigated. UCCA was then normalized by these factors respectively. On the basis of these results, an optimal factor was selected and applied to 29 MS subjects. An extremely strong correlation between UCCA and LECA was seen (r= .88, P < .01). The coefficient of variation (COV) of UCCA was reduced to 4.4% from 9.3% after correction by LECA. The normalized measurement of UCCA correlated better with the expanded disability status scale (EDSS) than the absolute measurement especially in relapsing-remitting multiple sclerosis patients. Moreover, more spinal cord atrophy was identified in corrected data than uncorrected data. Our findings suggest that LECA is an optimal correction factor for UCCA and normalized UCCA is preferable to absolute measurement in cross-sectional study.


Added-value biomolecules' production from cocoa pod husks: A review.
Cocoa beans are produced through on-farm processing where residual biomass is discarded, including cocoa pod husks (CPH), cocoa bean shells and cocoa sweatings. CPH represents about 80% of these residues that are generated during the initial cocoa bean processing steps and their disposal occupies large areas, causing social and environmental concerns. In the last decades, the lignocellulosic composition of CPH has attracted the attention of the scientific and productive sector. Recently, some studies have reported the use of CPH in the production of medium to high value-added molecules, with potential applications in food and feed, agriculture, bioenergy, and other segments. This review presents biotechnological approaches and processes for the exploitation of CPH, including pre-treatment methods for the production of different biomolecules. Great perspectives and innovations were found concerning CPH exploitation and valorisation, but still more efforts are needed to valorise this potential feedstock and give support to producers in-development countries.