Selection of Riboflavin Overproducing Strains of Lactic Acid Bacteria and Riboflavin Direct Quantification by Fluorescence.
Riboflavin (vitamin B2) is a vitamin of the B group involved in essential biological pathways, including redox reactions and the electron transport chain. Some lactic acid bacteria (LAB) can synthesize riboflavin and this capability is strain-dependent. In the last years, a growing interest has focused on the selection of riboflavin-overproducing food-grade LAB for the vitamin biofortification of fermented foods, as well as for the formulation of innovative functional products.In this chapter we report fast and inexpensive techniques in order to (1) screen LAB isolates able to produce riboflavin from different matrices, (2) select spontaneous roseoflavin-resistant riboflavin overproducing strains, and (3) quantify vitamin B2 in culture media by fluorescence detection.These protocols could be useful to select new overproducing strains and/or species from different ecological niches, as well as to optimize the conditions for vitamin bioproduction.


Augmenting hematoma-scavenging capacity of innate immune cells by CDNF reduces brain injury and promotes functional recovery after intracerebral hemorrhage.
During intracerebral hemorrhage (ICH), hematoma formation at the site of blood vessel damage results in local mechanical injury. Subsequently, erythrocytes lyse to release hemoglobin and heme, which act as neurotoxins and induce inflammation and secondary brain injury, resulting in severe neurological deficits. Accelerating hematoma resorption and mitigating hematoma-induced brain edema by modulating immune cells has potential as a novel therapeutic strategy for functional recovery after ICH. Here, we show that intracerebroventricular administration of recombinant human cerebral dopamine neurotrophic factor (rhCDNF) accelerates hemorrhagic lesion resolution, reduces peri-focal edema, and improves neurological outcomes in an animal model of collagenase-induced ICH. We demonstrate that CDNF acts on microglia/macrophages in the hemorrhagic striatum by promoting scavenger receptor expression, enhancing erythrophagocytosis and increasing anti-inflammatory mediators while suppressing the production of pro-inflammatory cytokines. Administration of rhCDNF results in upregulation of the Nrf2-HO-1 pathway, but alleviation of oxidative stress and unfolded protein responses in the perihematomal area. Finally, we demonstrate that intravenous delivery of rhCDNF has beneficial effects in an animal model of ICH and that systemic application promotes scavenging by the brain's myeloid cells for the treatment of ICH.


Gender differences in body fat content are present well before puberty.
To determine whether gender differences in body fat could be detected in prepubertal children using dual energy X-ray absorptiometry (DEXA), body composition was measured in 20 healthy boys aged 3-8 y matched for age, height and weight with 20 healthy girls. Although boys and girls did not differ in age, height, weight, body mass index (BMI) or bone mineral content, the boys had a lower percentage of body fat (13.5 +/- 5.1 vs 20.4 +/- 6.1%, P < 0.01), a lower fat mass (3.2 +/- 2.0 vs 4.9 +/- 3.1 kg, P < 0.01), and a higher bone-free lean tissue mass (18.6 +/- 4.3 vs 17.0 +/- 3.5 kg, P < 0.01) than the girls. Girls had approximately 50% more body fat than the boys. This is the first DEXA study to show that boys aged 3-8 y have less body fat than girls of similar age, height and weight. Thus, this technology demonstrates that significant gender differences in body composition are evident, well before the onset of puberty.


CRISPR-Based Transcriptional Activation in Drosophila.
Overexpression is one of the classical approaches to study pleiotropic functions of genes of interest. To achieve overexpression, we often increase the transcription by introducing genes on exogenous vectors or by using the CRISPR/dCas9-based transcriptional activation system. To date, the most efficient CRISPR/dCas9-based transcriptional activator is the Synergistic Activation Mediator (SAM) system whereby three different transcriptional activation domains are directly fused to dCas9 and MS2 phage Coat Protein (MCP), respectively, and the system in Drosophila is named flySAM. Here we describe the effective and convenient transcriptional activation system, flySAM, starting from vector construction, microinjection, and transgenic fly selection to the phenotypic analysis.


Salt overload damages the glycocalyx sodium barrier of vascular endothelium.
Sodium overload stiffens vascular endothelial cells in vitro and promotes arterial hypertension in vivo. The hypothesis was tested that the endothelial glycocalyx (eGC), a mesh of anionic biopolymers covering the surface of the endothelium, participates in the stiffening process. By using a mechanical nanosensor, mounted on an atomic force microscope, height (∼400 nm) and stiffness (∼0.25 pN/nm) of the eGC on the luminal endothelial surface of split-open human umbilical arteries were quantified. In presence of aldosterone, the increase of extracellular sodium concentration from 135 to 150 mM over 5 days (sodium overload) led the eGC shrink by ∼50% and stiffening by ∼130%. Quantitative eGC analyses reveal that sodium overload caused a reduction of heparan sulphate residues by 68% which lead to destabilization and collapse of the eGC. Sodium overload transformed the endothelial cells from a sodium release into a sodium-absorbing state. Spironolactone, a specific aldosterone antagonist, prevented these changes. We conclude that the endothelial glycocalyx serves as an effective buffer barrier for sodium. Damaged eGC facilitates sodium entry into the endothelial cells. This could explain endothelial dysfunction and arterial hypertension observed in sodium abuse.


Racial/ethnic equity in substance use treatment research: the way forward.
Opioid use and opioid-related overdose continue to rise among racial/ethnic minorities. Social determinants of health negatively impact these communities, possibly resulting in poorer treatment outcomes. Research is needed to investigate how to overcome the disproportionate and deleterious impact of social determinants of health on treatment entry, retention, drug use and related outcomes among racial/ethnic minorities. The current commentary provides recommendations that may help researchers respond more effectively to reducing health disparities in substance use treatment. We begin with recommendations of best research practices (e.g., ensuring adequate recruitment of racial/ethnic minorities in research, central components of valid analysis, and adequate methods for assessing effect sizes for racial/ethnic minorities). Then, we propose that more NIDA research focuses on issues disproportionately affecting racial/ethnic minorities. Next, techniques for increasing the number of underrepresented racial/ethnic treatment researchers are suggested. We then recommend methods for infusing racial/ethnic expertise onto funding decision panels. This commentary ends with a case study that features NIDA's National Drug Abuse Treatment Clinical Trials Network (CTN). The proposed recommendations can serve as guidelines for substance use research funders to promote research that has the potential to reduce racial/ethnic disparities in substance use treatment and to increase training opportunities for racial/ethnic minority researchers.


Performance-enhancing drugs: understanding the risks.
To help clinicians understand the risks associated with performance-enhancing drugs, this overview covers prohibited lists of substances and methods, therapeutic use exemptions, the legitimate indications and adverse effects, including for megadose and polypharmacy doping of stimulants, anabolic steroids, erythropoiesis-stimulating agents, and growth hormone and ways in which physicians or patients risk committing anti-doping rule violations inadvertently.


Human papillomavirus-Inequalities in disease prevention and the impact on racial, ethnic, sexual, and gender minorities.
HPV, a sexually transmitted viral infection, is the etiological agent of significant dermatologic disease including benign anogenital warts and invasive cancers. Sexual and gender minority individuals are particularly vulnerable to HPV-associated disease due to reduced vaccination rates in these cohorts, low awareness of HPV, lack of provider recommendation, and inadequate consensus guidelines on screening and prevention in these individuals. A targeted approach is needed with regards to vaccination in all children -especially those from racial, ethnic, sexual, and gender minorities; provider recommendation, especially from pediatric dermatologists, is crucial. Effort must also be made to use transgender and non-binary affirming language as dividing vaccination programs by anatomic sex and sexuality reinforces problematic notions of gender identity and sexuality, isolating the most vulnerable.


Experimental myonecrosis induced by the venoms of South American Micrurus (coral snakes).
Venoms from 11 taxa of Micrurus (coral snakes) from Brazil and Colombia were tested for myotoxic activity in mice. All venoms, except that of M. surinamensis, induced myotoxicity as judged by the increase in plasma creatine kinase levels and by histological analysis. Qualitatively, these venoms induced a similar necrotic pattern, although there were conspicuous quantitative differences between them.


Kinetics of Zn(2+)-induced brain type creatine kinase unfolding and aggregation.
We studied the effect of Zn(2+) on the folding and aggregation of brain creatine kinase (CK-BB). We developed a method to purify CK-BB from rabbit brain and conducted inhibition kinetics and unfolding studies of CK-BB. Zn(2+) conspicuously aggregated and osmolytes, such as glycine and proline, were able to suppress the formation of aggregates and protect the enzymatic activity against Zn(2+). These results suggest that Zn(2+) might act as a risk factor for CK-BB in the brain under certain conditions, and some osmolytes may help CK-BB to sustain the active state when Zn(2+) is present. Our study provides useful information regarding the effect of Zn(2+) on brain-derived metabolic enzymes, especially those that are putatively related to brain disease. Furthermore, our study suggests that although Zn(2+) may induce CK-BB inactivation and misfolding, the ability of some abundant proteins and osmolytes to chelate Zn(2+) nonspecifically may protect CK-BB and allow it to exist in the active form.


Radiographic patterns and viral studies in childhood pneumonia at various ages.
We aimed at evaluating the relationship between microbial etiology and chest radiograph appearance in various types of pneumonia. In a prospective study, the radiographic findings in 479 cases of acute pneumonia in children were compared with viral etiology and growth of potential bacterial pathogens in nasopharyngeal secretion. As the basis for viral etiology was most conclusive, the material was here classified according to the viral findings. The patients were divided into three age groups: 0-2, 3-5 and 6-15 years. The chest radiograms were analyzed blindly for the presence of hyperinflation and interstitial, alveolar and mixed interstitial-alveolar infiltrates. There was a statistically significant relationship between low age and occurrence of hyperinflation and interstitial infiltrates, and between high age and alveolar infiltrates. No unequivocal relationship was found between type of infiltrates or presence of atelectasis and proven viral etiology. We conclude that chest radiographs are not a useful indicator of microbial etiology in childhood pneumonia.


Two techniques for assessing postoperative voiding function, a randomized trial.
Post-operative voiding dysfunction is common after sling placement and is assessed with a voiding trial (VT) before discharge. We hypothesized that an operating room (OR) initiated retrograde VT (OR-fill) would decrease time to discharge compared with a post-anesthesia care unit (PACU) initiated retrograde VT (PACU-fill). This was a parallel non-blinded randomized trial, of women undergoing outpatient sling surgery at a university hospital. In the OR-fill group, fill was initiated after sling placement during cystoscopy by leaving fluid in the bladder. In the PACU-fill group, fill was initiated in the PACU. Pre-VT volumes were recorded and a PVR ≤ 100 mL defined a passed VT. The primary outcome was total PACU time, defined as arrival in PACU until discharge home. Thirty women were assigned to the OR-fill group and 29 to the PACU-fill group. Time from PACU arrival to VT was shorter in the OR-fill group (41.5 [31.0, 69.3] min vs 69.0 [44.0, 107.0] min, p = 0.03), but total PACU time in the groups was similar (125.0 [90.5, 180.5] min vs 131.5 [93.5, 178.0] min, p = 0.76). Bladder volume before VT was greater in the OR-fill group (557.3 ± 187.3 mL vs 433.0 ± 171.2 mL, p = 0.01). VT pass rates of the two groups were not significantly different (OR-fill 16.7% vs PACU-fill 24.1%, p = 0.48). OR-fill VT did not decrease total PACU time compared with PACU-fill VT. No overdistension was seen in either group; transient postoperative voiding dysfunction was common.


Boron concentration in water, sediment and different organisms around large borate deposits of Turkey.
Boron is an essential nutrient for plants and an essential element for many organisms, but can be toxic to aquatic and terrestrial organisms above certain concentrations. The aim of this research was to determine boron concentrations in water, sediment and biotic samples (Gammaridae spp.-Crustacea, Helix sp.-Gastropoda, Donax sp.-Bivalvia, Helobdella sp.-Hirudinae, Ephemeroptera nymph, Chrinomidae larvae, Tipulidae larvae-Insecta, Rana sp.-Amphibia, Natrix sp.-Serpentes, fish sample Leiscus cephalus (Linnaeus, 1758) and leaves of Salix sp.-Salicacea from Seydi Stream (Kirka-Eskişehir). Our results have shown that boron concentrations of the Seydi Stream water is higher than the Turkish Environmental Guidelines standard (>1 mg L(-1)) and in Europe (mean values typically below 0.6 mg L(-1)).


Pulsed UV-light inactivation of poliovirus and adenovirus.
To study the pulsed ultraviolet (UV) inactivation of poliovirus and adenovirus. Viral suspensions of 2 ml volume were exposed to varying numbers of polychromatic light pulses emitted from a xenon flashlamp. Ten pulses produced an approximately 4 log(10) reduction in poliovirus titre, and no infectious poliovirus remained after 25 pulses. With adenovirus, 10 pulses resulted in an approximately 1 log(10) reduction in infectivity. Adenovirus required 100 pulses to produce an approximately 3 log(10) reduction in infectivity, and 200 pulses to produce a greater than 4 log(10) reduction. Adenovirus was more resistant to pulsed UV treatment than poliovirus although both viruses showed susceptibility to the treatment. Pulsed UV-light treatment proved successful in the inactivation of poliovirus and adenovirus, and represents an alternative to continuous-wave UV treatment.


A Simple Prognostic Scoring System for Hepatocellular Carcinoma Treated with Selective Internal Radiation Therapy.
Selective internal radiation therapy (SIRT) is a local treatment option for patients with hepatocellular carcinoma (HCC). Its exact role next to other HCC therapies has yet to be defined. In order to identify patients most suitable for SIRT, a SIRT-specific prognostic score should be developed. A cohort of 72 SIRT patients treated at the University Hospital of Munich was retrospectively analyzed. The prognostic performance of 12 HCC staging systems and prognostic scores was assessed. Cox-regression analysis was used to identify independent prognostic factors, which formed the basis of the Munich-SIRT score (M-SIRT). All scores were ranked by calculating the c-Index and Akaike information criterion (AIC). External validation was performed in a cohort of 128 SIRT patients treated at the University Hospital of Pamplona, Spain. median overall survival was 13 months (95% confidence interval 9.9-21.9). AFP (p = 0.005; hazard ratio [HR] 2.38), albumin (p < 0.001; HR 5.87), and alkaline phosphatase (p < 0.001; HR 8.38) were identified as independent prognostic factors. M-SIRT comprises 3 prognostic groups with a median survival of 38.9, 14.6, and 7.7 months, respectively (I vs. II: p = 0.003, II vs. III: p < 0.001). AIC (318) and concordance index (0.711) ranked M-SIRT superior to the established HCC staging systems, and the score successfully passed external validation in an independent SIRT cohort (I vs. II: p = 0.03; II vs. III: p = 0.007). Therapy-specific prognostic scores can facilitate treatment decisions and prognostication for HCC patients. Considering its performance in 200 SIRT patients, M-SIRT is a promising prognostic tool for HCC patients evaluated for SIRT.


Source apportionment and human health risk of PAHs accumulated in edible marine organisms: A perspective of "source-organism-human".
Most PAHs produced by human activities can be absorbed and accumulated by edible organisms and pose a potential hazard to human health. However, the source apportionment and human health risk of PAHs accumulated in edible organisms remains largely unknown. Therefore, we conducted source analysis and health risk assessment based on the PAH concentrations in ten marine fish from coastal areas of Guangdong, China. Results showed that the pollution of PAHs in fish organisms was at "Minimally polluted" level, and that all marine fish had the ability to accumulate PAHs. Risk assessment indicated Carcinogenic risk of PAHs in four populations was at a "Cautionary risk" level, with urban children suffered the highest risk. Petroleum pollution, Coal and biomass combustion, and Marine transport emissions were identified as the main anthropogenic sources for PAHs in organisms, and Marine transport emissions accounted for the highest Carcinogenic risk. The Acceptable daily intake for all populations were far below their actual daily intake without causing "Cautionary risk". Our findings provide new insights into the source apportionment and health risk of PAHs from a "source-organism-human" perspective, and suggested that joint management of three anthropogenic sources would be an effective way to prevent the health risks of PAHs.


Cervical spine disorders in farm workers requiring neck extension actions.
Cervical extension is considered a risk factor for provoking radicular and spinal cord compromise. However, there have been no reports on the relation between extension strain (ES) and cervical spine disorders: degenerative cervical spine disorders, dynamic canal stenosis (DCS), and cervical myelopathy. We performed a cross-sectional study to investigate the relation. Orthopedic examinations were performed on 177 grape-growers (ES-exposed group) and 191 eggplant-growers (control group) between May and August 2000; and patients with degenerative cervical spine disorders, DCS, and cervical myelopathy were identified in the two groups. ES, degenerative cervical spine disorders, DCS, and cervical myelopathy were regarded as exposure variables; and age, sex, and working years were regarded as confounders. In cases where the subject was exposed to each exposure variable, multivariate-adjusted odds ratios to degenerative cervical spine disorders, DCS, and cervical myelopathy and multivariate-adjusted odds ratios regarding cervical myelopathy as the dependent variable were calculated by unconditional logistic regression analysis. Multivariate unconditional logistic regression analysis showed a significant odds ratio of ES to degenerative cervical spine disorders [2.72, 95% confidence interval (CI) 1.62-4.56]. Multivariate unconditional logistic regression analysis showed significant odds ratios for DCS (4.50, 95% CI 2.03-9.96) and age (1.07, 95% CI 1.01-1.14) regarding cervical myelopathy as the dependent variable. These findings suggested that ES of the cervical spine is a risk factor for degenerative cervical spine disorders, and DCS and the aging process are risk factors for cervical myelopathy.


Periprostatic innervation: New issues based on segmental analysis of 10 human cadaver pelvic blocs.
The exact paths of periprostatic nerves have been under debate over the last decades. In the present study, the topographic distribution of nerves around the prostate and their relative distances from the prostatic capsule were analyzed in male cadaver visceral blocs. The pelvic organs from ten fresh male cadavers were removed and serial sectioned en bloc for histological investigation. The macroslices was divided into four sectors. Each sector was centrally covered with a raster dividing each sector in three subsectors numbered clockwise. The prostatic capsule was identified, and distances of 2.5 and 5 mm from the prostate were demarked with lines. We quantified the number of nerve fibers present in each subsector of each slide and recorded their position relative to the prostatic capsule. In general, the topographic analysis revealed that the majority of nerves were identified in sectors 4 through 9, corresponding to the posterolateral and posterior surfaces of the prostate gland. At the prostate base, the majority of nerves were found at the posterolateral and posterior surfaces of the gland. Within the mid-region of the prostate, the same topographic distribution pattern was observed, but the nerve fibers were closer to the prostatic capsule. At the apical region, the percentage of nerve fibers identified in the anterior region was higher, despite their major concetration in the posterior surface. The nerves identified at the apex were mainly located up to 2.5 mm from the prostate. This proximity to the prostate was specifically observed in the anterolateral and anterior sectors. In the craniocaudal sense, the percentage of nerves identified between 2.5 and 5 mm from the prostatic capsule remained constant. A significant number of nerve fibers were present in the anterior and anterolateral positions, especially at the apex. The anterior nerves were closer to the prostate. This proximity suggests that the anterior nerves may participate in local physiology and that the cavernous nerves are probably formed by the posterior nerve fibers. It is likely that the safe distance of 2.5 mm from all surfaces of the prostate may be related to cavernous fiber preservation.


Rapid quantification of royal jelly quality by mid-infrared spectroscopy coupled with backpropagation neural network.
Royal jelly is rich in nutrients but its quality is greatly affected by storage conditions. To determine the quality of royal jelly accurately and quickly, a qualitative discrimination model was established based on the fusion of conventional parameters and mid-infrared spectrum, using support vector machine. The prediction models for three representative quality parameters were developed by the backpropagation neural network with various algorithms. The results demonstrated that the recognition rate of the multi-source information fusion model was increased to 100% when compared with that of the spectral data preprocessed by Savitzky-golay smoothing (95.83%). The mean square errors of the constructed model for moisture, water-soluble protein, and total sugar were 0.0032, 0.0058, and 0.0069, respectively. The constructed model had an ensured accuracy for the calibration set, with the correlation coefficient of prediction maintained at 0.9353, 0.9533, and 0.9563, which could meet the requirement of non-destructive rapid detection of royal jelly quality.


Multipathogen oligonucleotide microarray for environmental and biodefense applications.
Food-borne pathogens are a major health problem. The large and diverse number of microbial pathogens and their virulence factors has fueled interest in technologies capable of detecting multiple pathogens and multiple virulence factors simultaneously. Some of these pathogens and their toxins have potential use as bioweapons. DNA microarray technology allows the simultaneous analysis of thousands of sequences of DNA in a relatively short time, making it appropriate for biodefense and for public health uses. This paper describes methods for using DNA microarrays to detect and analyze microbial pathogens. The FDA-1 microarray was developed for the simultaneous detection of several food-borne pathogens and their virulence factors including Listeria spp., Campylobacter spp., Staphylococcus aureus enterotoxin genes and Clostridium perfringens toxin genes. Three elements were incorporated to increase confidence in the microarray detection system: redundancy of genes, redundancy of oligonucleotide probes (oligoprobes) for a specific gene, and quality control oligoprobes to monitor array spotting and target DNA hybridization. These elements enhance the reliability of detection and reduce the chance of erroneous results due to the genetic variability of microbes or technical problems with the microarray. The results presented demonstrate the potential of oligonucleotide microarrays for detection of environmental and biodefense relevant microbial pathogens.


Single-stranded DNA of 5'-upstream region of the rolC gene interacts with nuclear proteins of carrot cell cultures.
Using the gel retardation assay, proteins of carrot cells capable of binding to a single-stranded DNA of 5'-upstream region of the rolC gene were found. From competition experiments, these DNA-protein interactions were specific to single-stranded nucleotide sequence of Ava S fragment (from -94 bp to +23 bp relative to the transcription initiation site). Methylation interference experiments showed that G residue at the position of -41 bases on the bottom strand was important for DNA-protein binding. This residue was located between CAAT box and TATAA box. Such specific interaction between single-stranded DNA and nuclear proteins may play a role in transcription by RNA polymerase II.


Toxicity of dichloropropanols in rat hepatocyte cultues.
Exposure of humans to dichloropropanols has been shown to result in fulminant hepatic necrosis. These compounds have also been shown to be hepatotoxic in rats. In this study, 1,3-dichloropropanol, but not 2,3-dichloropropanol, was shown to be toxic to 24 h cultures of rat hepatocytes. The toxicity was inhibited by pre-treatment of cultures with a cytochrome P450 inhibitor and enhanced by prior depletion of cellular glutathione. In addition, at equimolar concentrations both isomers were shown to deplete glutathione, although the extent of depletion was greater with the 1,3-isomer. 1,3-Dichloropropanol also depleted ATP and reduced the mitochondrial membrane potential. The effects on ATP, glutathione and membrane potential could be inhibited by the cytochrome P450 inhibitor. It is concluded that the toxicity of 1,3-dichloropropanol is mediated by cytochrome P450 and involves depletion of glutathione and loss of mitochondrial function.


A novel coumarin-derived acylhydrazone Schiff base gelator for synthesis of organogels and identification of Fe3.
A novel coumarin-derived acylhydrazone Schiff base fluorescent organogel (G1) was designed and synthesized. Gelator G1 can form stable organogels in isopropanol, tert-amyl alcohol, n-butanol and phenylamine. The organogel could be converted to solution by heating and the solution could be restored to gel state by cooling. The self-assemble mechanism of G1 was investigated by XRD, FT-IR and SEM techniques. The results indicated the intermolecular hydrogen bonding, Van der Waals interaction and π-π stacking are the forces for the self-assembly of the gelator to form the organogel. The optical properties of the compound were studied by UV-visible spectroscopy and fluorescence spectra. Further study presented that gelator G1 could selectively and sensitively response to Fe3+ only among tested cations. Beside the above functions, the organic gel factor G1 could also response to irradiation, heating and shaking, thus endowing the organogel with multi stimulus responsive properties.


The eggshell ofDrosophlla melanogaster : VII. Formation of the micropylar canal and the role of the paracrystalline structure.
The micropylar apparatus (MA) inDrosophila melanogaster (Diptera: Drosophilidae) is a 25 μm long protrusion on the anterior pole of the eggshell, and it contains the 0.8 μm wide micropylar canal through which the spermatozoon penetrates the eggshell. The canal terminates in the paracrystalline structure thus forming the "pocket". The MA is secreted during oogenesis by the micropylar follicle cells (border cells and peripheral cells). Morphogenesis of the micropylar canal starts before stage 11A, when two of the border cells form two thin extensions containing microfilaments and penetrate the paracrystalline structure. Microtubules found at the base of the extensions participate in the formation of two projections. Adjacent follicle cells secrete the chorionic part of the MA, whilst the two projections elongate and twist during stages 12 and 13. Microtubules run parallel to the long axis of the projections and probably are related to cellular elongation during the formation of the projections. The paracrystalline structure is composed of lamellae having a periodicity of about 50 nm, which is maintained possibly due to the lateral fibrils connecting the lamellae. These lamellae consist of small fragments secreted during stages 10-11 by the border cells. The paracrystalline structure and the spongy vitelline membrane are condensed after egg maturation.


Expression and functional analysis of the lysine decarboxylase and copper amine oxidase genes from the endophytic fungus Colletotrichum gloeosporioides ES026.
Huperzine A (HupA) isolated from Huperzia serrata is an important compound used to treat Alzheimer's disease (AD). Recently, HupA was reported in various endophytic fungi, with Colletotrichum gloeosporioides ES026 previously isolated from H. serrata shown to produce HupA. In this study, we performed next-generation sequencing and de novo RNA sequencing of C. gloeosporioides ES026 to elucidate the molecular functions, biological processes, and biochemical pathways of these unique sequences. Gene ontology and Kyoto Encyclopedia of Genes and Genomes assignments allowed annotation of lysine decarboxylase (LDC) and copper amine oxidase (CAO) for their conversion of L-lysine to 5-aminopentanal during HupA biosynthesis. Additionally, we constructed a stable, high-yielding HupA-expression system resulting from the overexpression of CgLDC and CgCAO from the HupA-producing endophytic fungus C. gloeosporioides ES026 in Escherichia coli. Quantitative reverse transcription polymerase chain reaction analysis confirmed CgLDC and CgCAO expression, and quantitative determination of HupA levels was assessed by liquid chromatography high-resolution mass spectrometry, which revealed that elevated expression of CgLDC and CgCAO produced higher yields of HupA than those derived from C. gloeosporioides ES026. These results revealed CgLDC and CgCAO involvement in HupA biosynthesis and their key role in regulating HupA content in C. gloeosporioides ES026.


Rapid bacteria-detection platform based on magnetophoretic concentration, dielectrophoretic separation, and impedimetric detection.
Most biosensors employ small sample quantities (less than 100 μL) for bacteria detection, thereby resulting in inaccurate low-concentration measurements. Detection performed using small sample volumes with low bacteria concentration may produce false-negative results. Therefore, sample pretreatment plays a critical role in accurate bacteria detection. This paper presents an impedimetric bacteria-detection sensor integrated with bacteria concentration and separation devices for rapid bacteria detection. Post conjugation using magnetic particles (MPs), the MP-conjugated bacteria (MP/Bac) are concentrated via magnetophoresis by a factor exceeding 100. In addition, MP/Bac are separated from MPs via dielectrophoresis to prevent occurrence of signal errors caused by MPs not conjugated with bacteria. Subsequently, concentrated MP/Bac are captured on a sensor electrode, and bacteria concentration is detected by measuring signal changes caused by the impedance difference between bacteria and the medium. The performance of the proposed bacteria-detection device was evaluated using a 5-mL homogenized cabbage sample injected with Staphylococcus aureus at 30 mL/h flow rate. The observed signal change was measured for 10 min using a sample with a concentration of 5-5 × 103 CFU/mL and was found to be approximately 0.34 mV at 50 CFU/mL; the limit of detection was 36 CFU/mL. These results confirm that the proposed device can detect low bacteria concentrations in food samples.


Adaptive optimal sound for vocal communication in tunnels of a subterranean mammal (Spalax ehrenbergi).
The adaptive value of sound signal characteristics for transmission in the underground tunnel ecotope was tested using tunnels of the solitary territorial subterranean mole rats. We analyzed the propagation of synthetic calls with various frequencies through natural tunnels along different distances. Here we present evidence that sound propagation proved efficient only across short distances (a few meters). The least attenuation of sounds occurred at low frequencies. The 440 Hz sound was transmitted better than the lower (220 Hz) or higher (880, 1760, 3520 Hz) tested frequencies. These characteristics matched perfectly with the mole rat features of vocalization and hearing, thus reflecting the operation of natural selection for adaptive vocal communication in the underground tunnel ecotope.


Arsenic and trace metal concentrations in different vegetable types and assessment of health risks from their consumption.
The levels of 12 trace metal (loid)s (TMs) in 10 vegetable types including leafy vegetables (purslane, purple basil and parsley) and fruiting vegetables (pepper, tomato, eggplant, cucumber, zucchini, green bean and melon) and in maize grown in Malatya province (Turkey) were investigated and non-carcinogenic health risks from consumption of these crops were assessed. The levels of TMs were measured by inductively coupled plasma-optical emission spectrometry. The mean levels of Ni, Cu, Cd, Cr, As and Zn in all crops were below maximum permissible concentrations (MPCs), while those of Pb in pepper (0.109 mg/kg fw), eggplant (0.103 mg/kg fw) and green bean (0.177 mg/kg fw) slightly exceeded MPCs (0.1 mg/kg fw). Leafy vegetables had relatively higher concentrations of Al, As, Ba, Cr, Cu, Fe and Mn compared to fruiting vegetables and maize. The estimated daily intake value of each TM estimated for each crop was found to be below the tolerable daily intake value. The target hazard quotients of all TMs in all crops did not exceed the acceptable non-carcinogenic risk level. However, hazard index (HI) value (1.57) in tomato was found to be above the threshold value of 1, indicating non-carcinogenic risks for consumers due to the intake of combined TMs in tomato. The THQ values of As, Co and Pb contributed 46.4%, 24.5% and 16.4% of the HI value of tomato, respectively. High daily consumption amount of tomato likely resulted in high HI value. The findings obtained in this study reveal that even if the levels of TMs in vegetables are safe, more attention should be paid to non-carcinogenic risks associated with TMs as a result of high vegetable intake.


Longitudinal evaluation of posterior corneal power by anterior segment optical coherence tomography 18 months following photorefractive keratectomy.
To investigate the change in anterior, posterior, and net corneal power more than 18 months after photorefractive keratectomy (PRK) by RTVue anterior segment optical coherence tomography (OCT). Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran. Prospective observational study. PRK was performed using Technolas TENEO 317 laser platform. Anterior, posterior, and net corneal power was measured by the RTVue-XR anterior segment OCT system with the Pachymetry + Cpwr scan pattern at baseline and months 1, 3, 6, 12, 18, and more than 18 (up to 27) after PRK. Three hundred twenty-six eyes of 163 patients (61 men [37.4%], 102 women [62.6%]; mean age 29.71 years; range 18.5 to 46.5 years) were enrolled in this study. Mean preoperative spherical equivalent was -3.15 ± 1.50 diopter (D) (range -8.37 to -0.62 D). The mean change in net corneal power was 3.052 D, 3.281 D, 3.324 D, 3.114 D, 3.446 D, and 3.972 D at months 1, 3, 6, 12, 18, and more than 18 postoperatively compared with baseline, respectively (P < .001 for all comparisons). The mean change in posterior corneal power at postoperative visits compared with baseline was not statistically significant (P > .1) except for 1 month postoperatively, which increased by 0.13 D (P < .001). Changes in posterior corneal power were not correlated with any of the preoperative clinical and Scheimpflug variables. Posterior corneal power did not change for more than 18 months after PRK, except for an early small increase at 1 month postoperatively.


[Vascularized periosteum and bone regeneration].
The osteogenic potential of periosteum is widely recognized. During development, it plays a prominent role in the radial growth of long bones. Similarly, it has a key role in the consolidation of fractures. The physiological function of periosteum in the healthy, mature skeleton remains relatively subtle; however, its detachment from the bone surface reactivates its potential for fibrogenic and osteochondrogenic regeneration. This discreet anatomical structure is actually a reservoir of mesenchymal progenitor cells capable of proliferating and differentiating, by reinitializing cellular and molecular cascades of embryogenesis in mesenchymal tissues. However, given the hitherto limited knowledge of the quantitative potential of periosteum and of the pathways regulating tissue differentiation during regeneration, human applications have remained anecdotal. The findings of several in vivo and in vitro experiments indicate that the maintenance of the periosteum's vascularization stimulates its quantitative potential. The structural organization of the regenerated material in vivo is governed by locoregional biological and mechanical regulatory mechanisms that serve to make it capable of performing its new functions. The increasing awareness of periosteum's potential is stimulating active research in the fields of cellular biology and tissue engineering. The demonstration of its regenerative potential in animals gives reason to believe that strips of vascularized periosteum could become part of the developing armamentarium of regenerative medicine.


[The characteristics of treating odontogenic osteomyelitis against a background of malaria in the population of the Republic of Burundi].
All the patients were administered preoperative antimalarial therapy, antibiotics and sulfanilamides, and agents maintaining the defense factors, as well as detoxifying and desensitizing therapy. Rational nutrition was prescribed. All these measures helped liquidate the inflammation foci not only in the bones, but in the soft tissues of the paramaxillary area as well.


Preschool vision screening: a prospective comparative evaluation.
We have reviewed the results of a pilot study of preschool screening by orthoptists for vision defects which was introduced in Newcastle in 1987. We have compared the visual outcomes, at age 7 years, of children who were screened at age 3 years by either orthoptists, health visitors or general practitioners in three matched, geographically defined cohorts. Manifest, large angle strabismus presented at the same age, and in roughly equal numbers in each cohort. Orthoptic screening detected many more cases of amblyopia associated with microtropia and anisometropia, but the b overall amblyopia prevalence at age 7 years was similar in each cohort. This study does not provide evidence to support the nationwide introduction of primary orthoptic preschool vision screening, and highlights the need for a prospective treatment trial of amblyopia associated with microtropia and anisometropia.


[The clinical significance of the sulfhydryl groups in the peripheral blood lymphocytes of patients with thyrotoxic goiter].
A study of 42 patients with thyrotoxic goiter revealed a reduction of SH groups in the peripheral blood lymphocytes. The reduction depended on the severity of the pathological process. Changes of the SH groups are an informative criterion of the severity of the disease and treatment efficacy.


Treatment strategies for diabetic patients suffering from erectile dysfunction: an update.
Erectile dysfunction (ED) is one of the most common complications of diabetes. The complex pathophysiology of this disease makes it difficult to create clinical treatments tailored specifically for diabetic patients. This article will provide an update of both medical and surgical treatment approaches for diabetic ED, as well as discuss established treatments that are the cornerstone of this field. Currently, PDE type 5 inhibitors represent the first-line treatment for all patients with ED. In the interim since this article's first publication several new PDE5 inhibitors have come to market. Several others that were previously on the market are now available for daily dosing. Analysis of both testosterone level and hemoglobin A1c is an essential part of treatment. Intracavernosal injections and vacuum constriction devices remain safe, and are highly effective treatment options. The implantable penile prosthesis remains a safe, and definitive treatment modality for those patients with refractory ED. Over the coming years as the understanding of the underlying causes of diabetic ED continues, the development of new, noninvasive, but effective medical treatments will lead to even more options for the treatment of this difficult disease.


Development and Application of an Interdisciplinary Rapid Message Testing Model for COVID-19 in North Carolina.
From the onset of the COVID-19 pandemic, public health officials have sought to develop evidence-based messages to reduce COVID-19 transmission by communicating key information to media outlets and the public. We describe the development of an interdisciplinary rapid message testing model to quickly create, test, and share messages with public health officials for use in health campaigns and policy briefings. An interdisciplinary research team from the University of North Carolina at Chapel Hill assembled in March 2020 to assist the state health department in developing evidence-based messages to influence social distancing behaviors in the state. We developed and iteratively executed a rapid message testing model; the components of the 4-step model were message creation, survey development, survey administration, and analysis and presentation to health department officials. The model was executed 4 times, each during a 7-day period in April and May, and each subsequent survey included new phrasing and/or messaging informed by the previous week's survey. A total of 917 adults from North Carolina participated in the 4 surveys. Survey participants rated messages focused on protecting oneself and others higher than messages focused on norms and fear-based approaches. Pairing behaviors with motivations increased participants' desire to social distance across all themes and subgroups. For example, adding "Protect your grandmother, your neighbor with cancer, and your best friend with asthma," to messaging received a 0.9-point higher score than the base message, "Stay 6 feet apart from others when out in public." Our model to promote social distancing in North Carolina during the COVID-19 pandemic can be used for rapid, iterative message testing during public health emergencies.


Social support and sleep quality in patients with stroke: The mediating roles of depression and anxiety symptoms.
Research has demonstrated that higher social support is associated with better psychological health, quality of life, cognition, activities of daily living and social participation, but the relationship between social support and sleep quality remains unknown. This study aimed to assess the incidence of poor sleep quality, clarify the relationship between social support and sleep quality amongst stroke patients and determine whether anxiety and depression symptoms mediate this relationship. We conducted a quantitative, cross-sectional study involving 238 patients with stroke (median age of 61 [range 29-87] years, 68.1% male) recruited from a comprehensive tertiary care hospital between September 2019 and January 2020. A self-administered, structured questionnaire was used for the survey. The mediating effect of anxiety and depression symptoms was assessed using the bootstrap method via Model 4 (parallel mediation) of the SPSS PROCESS macro. Results showed that the incidence of poor sleep quality amongst stroke patients was 65%. Mediation analysis showed that social support exerted significant direct effects on sleep quality, and anxiety and depression symptoms mediated the relationship between social support and sleep quality. Measures should be taken to enhance social support to improve the sleep quality of stroke patients.


DNA sequence analysis of Tn10 insertions: origin and role of 9 bp flanking repetitions during Tn10 translocation.
The sequences of insertions of the translocatable tetracycline-resistance element Tn10 into the repressor (cl) gene of bacteriophage lambda have been analyzed. Each insertion contains the same discrete set of Tn10 sequences flanked by a direct repetition of a 9 bp cl-gene sequence. The flanking repititions are generated by duplication of information present only in the target DNA molecule rather than by a Campbell-type recombination event between one 9 bp sequence on the target DNA and a second one provided on the incoming element. The repetitions do not contain genetic or structural information important for translocation. A genetically constructed Tn10 insertion which lacks flanking repetitions is fully functional in translocation to a new position. Tn10 insertions cluster at preferred positions along a target DNA (Kleckner et al., 1979). Sequence analysis shows that four independently isolated cl::Tn10 insertions occur at identical positions in the cl gene. We speculate that homology between Tn10 and its target, at some distance from the site of the actual recombination event, could be relevant to the preference of Tn10 for particular insertion sites.


Inactivation of the infralimbic cortex decreases discriminative stimulus-controlled relapse to cocaine seeking in rats.
Persistent susceptibility to cue-induced relapse is a cardinal feature of addiction. Discriminative stimuli (DSs) are one type of drug-associated cue that signal drug availability (DS+) or unavailability (DS-) and control drug seeking prior to relapse. We previously established a trial-based procedure in rats to isolate DSs from context, conditioned stimuli, and other drug-associated cues during cocaine self-administration and demonstrated DS-controlled cocaine seeking up to 300 abstinence days. The behavioral and neural mechanisms underlying trial-based DS-control of drug seeking have rarely been investigated. Here we show that following discrimination training in our trial-based procedure, the DS+ and DS- independently control the expression and suppression of cocaine seeking during abstinence. Using microinjections of GABAA + GABAB receptor agonists (muscimol + baclofen) in medial prefrontal cortex, we report that infralimbic, but not prelimbic, subregion of medial prefrontal cortex is critical to persistent DS-controlled relapse to cocaine seeking after prolonged abstinence, but not DS-guided discriminated cocaine seeking or DS-controlled cocaine self-admininstration. Finally, using ex vivo whole-cell recordings from pyramidal neurons in the medial prefrontal cortex, we demonstrate that the disruption of DS-controlled cocaine seeking following infralimbic cortex microinjections of muscimol+baclofen is likely a result of suppression of synaptic transmission in the region via a presynaptic mechanism of action.


Psychological correlates of male child and adolescent sexual experiences with adults: a review of the nonclinical literature.
Researchers have generally neglected sexual experiences of boys with adults, assumed them to be the same as those of girls, or tried to understand them by referring to clinical research while ignoring nonclinical research. A review of nonclinical research allows a more complete understanding of boys' sexual experiences with adults and the outcomes and correlates of those experiences. Research with nonclinical samples reveals a broad range of reactions, with most reactions being either neutral or positive. Clinical samples reveal a narrower, primarily negative, set of reactions. Comparison of the reactions of boys and girls shows that reactions and outcomes for boys are more likely to be neutral or positive. Moderator variables, including presence of force, perceptions of consent, and relationship to the adult, also relate to outcomes. Incestuous contacts and those involving force or threats are most likely to be negative. Problems in this field of research include broad and vague definitions of "abuse" and conflation of value judgments with harm. Effects of boys' early sexual experiences with older persons in general cannot be accurately inferred from clinical research alone or from girls' experiences.


Control of red cell urea and water permeability by sulfhydryl reagents.
The binding constant for pCMBS (p-chloromercuribenzenesulfonate) inhibition of human red cell water transport has been determined to be 160 +/- 30 microM and that for urea transport inhibition to be 0.09 +/- 0.06 microM, indicating that there are separate sites for the two inhibition processes. The reaction kinetics show that both processes consist of a bimolecular association between pCMBS and the membrane site followed by a conformational change. Both processes are very slow and the on rate constant for the water inhibition process is about 10(5) times slower than usual for inhibitor binding to membrane transport proteins. pCMBS binding to the water transport inhibition site can be reversed by cysteine while that to the urea transport inhibition site can not be reversed. The specific stilbene anion exchange inhibitor, DBDS (4,4'-dibenzamidostilbene-2,2'-disulfonate) causes a significant change in the time-course of pCMBS inhibition of water transport, consistent with a linkage between anion exchange and water transport. Consideration of available sulfhydryl groups on band 3 suggests that the urea transport inhibition site is on band 3, but is not a sulfhydryl group, and that, if the water transport inhibition site is a sulfhydryl group, it is located on another protein complexed to band 3, possibly band 4.5.


Lack of effect of phenobarbital on hepatocellular carcinogenesis initiated by N-nitrosodiethylamine or methylazoxymethanol acetate in male Syrian golden hamsters.
Subchronic toxicity and long-term tumor-promoting effects of phenobarbital (PB) were investigated in male Syrian golden hamsters. In subchronic studies, PB was administered in drinking water to 5-week-old male hamsters for periods of 8 or 16 weeks at dosage levels of 250, 500, or 1000 ppm. No significant change in the ratio of liver weight to body weight was observed at 8 weeks; however, at 16 weeks there was a dose-dependent increase in the ratio of liver weight to body weight and a significant decrease in body weight gain among animals that received PB at 1000 ppm. The effect of PB on hepatic cytochrome P-450 and P-450-dependent aminopyrine N-demethylase activity was compared in male Syrian golden hamsters, F-344/NCr rats, and B6C3F1 mice. PB enhanced cytochrome P-450 activity in all three species; however, a significant increase (p less than 0.05) in aminopyrine N-demethylase activity was observed only in rats and mice. Potentially preneoplastic hepatocellular hyperplastic foci and hepatocellular neoplasms were studied in weanling male Syrian golden hamsters that received a single ip injection of either 100 mg N-nitrosodiethylamine (DEN)/kg body wt or 20 mg methylazoxymethanol acetate (MAM)/kg body wt at 5 weeks of age, followed by administration of 500 ppm PB in drinking water that began 2 weeks after the carcinogen injection and continued to 69 weeks of age. Groups of hamsters were killed at 25, 52, and 69 weeks of age; portions of liver and other organs with gross lesions were fixed in Formalin and examined histologically. MAM was a more potent hepatocarcinogen than DEN in male Syrian golden hamsters. PB failed to promote the development of either preneoplastic hepatocellular foci or hepatocellular neoplasms (adenomas or carcinomas) in either DEN- or MAM-initiated hamsters. Also, PB had no effect on the development of nonhepatic lesions occurring either spontaneously or induced by DEN or MAM in these animals.


Nanospan, an alternatively spliced isoform of sarcospan, localizes to the sarcoplasmic reticulum in skeletal muscle and is absent in limb girdle muscular dystrophy 2F.
Sarcospan (SSPN) is a transmembrane protein that interacts with the sarcoglycans (SGs) to form a tight subcomplex within the dystrophin-glycoprotein complex that spans the sarcolemma and interacts with laminin in the extracellular matrix. Overexpression of SSPN ameliorates Duchenne muscular dystrophy in murine models. Standard cloning approaches were used to identify nanospan, and nanospan-specific polyclonal antibodies were generated and validated. Biochemical isolation of skeletal muscle membranes and two-photon laser scanning microscopy were used to analyze nanospan localization in muscle from multiple murine models. Duchenne muscular dystrophy biopsies were analyzed by immunoblot analysis of protein lysates as well as indirect immunofluorescence analysis of muscle cryosections. Nanospan is an alternatively spliced isoform of sarcospan. While SSPN has four transmembrane domains and is a core component of the sarcolemmal dystrophin-glycoprotein complex, nanospan is a type II transmembrane protein that does not associate with the dystrophin-glycoprotein complex. We demonstrate that nanospan is enriched in the sarcoplasmic reticulum (SR) fractions and is not present in the T-tubules. SR fractions contain membranes from three distinct structural regions: a region flanking the T-tubules (triadic SR), a SR region across the Z-line (ZSR), and a longitudinal SR region across the M-line (LSR). Analysis of isolated murine muscles reveals that nanospan is mostly associated with the ZSR and triadic SR, and only minimally with the LSR. Furthermore, nanospan is absent from the SR of δ-SG-null (Sgcd-/-) skeletal muscle, a murine model for limb girdle muscular dystrophy 2F. Analysis of skeletal muscle biopsies from Duchenne muscular dystrophy patients reveals that nanospan is preferentially expressed in type I (slow) fibers in both control and Duchenne samples. Furthermore, nanospan is significantly reduced in Duchenne biopsies. Alternative splicing of proteins from the SG-SSPN complex produces δ-SG3, microspan, and nanospan that localize to the ZSR and the triadic SR, where they may play a role in regulating resting calcium levels as supported by previous studies (Estrada et al., Biochem Biophys Res Commun 340:865-71, 2006). Thus, alternative splicing of SSPN mRNA generates three protein isoforms (SSPN, microspan, and nanospan) that differ in the number of transmembrane domains affecting subcellular membrane association into distinct protein complexes.


Placental macrophage (Hofbauer cell) polarization is independent of maternal allergen-sensitization and presence of chorioamnionitis.
Macrophages can polarize in which M1/classically activated and M2/alternatively activated macrophages are considered to be the extremes. M1 macrophages are involved in inflammatory reactions, while M2 macrophages are suggested to be involved in homeostasis, parasite killing, tumor promotion, tissue remodeling and in allergic reactions. We hypothesized that polarization of placental macrophages (Hofbauer cells) is influenced by the allergen-sensitization status of the mother and/or the presence of chorioamnionitis, a placental inflammation. This Hofbauer cell polarization might be associated to the intrauterine environment and influence the risk of allergy development for the child. Therefore we aimed to determine the polarization status of Hofbauer cells in health and disease. We determined the expression of CD68, CX3CR1, IL-7R, DC-SIGN/CD209 and CD163 in placentas of sensitized versus non-sensitized mothers (n = 17), and placentas with or without histological chorioamnionitis (n = 10) by means of immunohistochemical analysis and quantitative real-time PCR (qPCR). Protein expression of the M1 markers (CX3CR1, IL-7R and CCR7) could not be detected in any of the analyzed samples while the M2 markers (DC-SIGN, CD163 and mannose receptor/CD206) were readily detected. Significant differences between non-sensitized versus sensitized mothers and uncomplicated versus chorioamnionitis complicated pregnancies were not detected at protein or at mRNA expression level. These results suggest that Hofbauer cells have an M2 phenotype, and that their polarization is not affected by maternal allergen-sensitization or by presence of chorioamnionitis.


Renal tubular dysfunction in methylmalonic acidaemia.
Renal tubular function was assessed in seven patients with methylmalonic acidaemia not responsive to vitamin B12. Five patients failed to concentrate their urine normally and in these patients the glomerular filtration rate was also reduced. Fractional excretion of sodium was increased in four patients, fractional excretion of potassium in one patient and in three there was a decreased tubular reabsorption of phosphate. Although possibly representing primary tubular damage these findings were thought to be consistent with adaptive changes secondary to the reduced glomerular filtration rate. Two patients had evidence of a defect of urinary acidification and several had a degree of hyporeninaemic hypoaldosteronism suggesting type 4 renal tubular acidosis. In one patient with a mild variant no renal disease was detected. Decreased renal function and tubular abnormalities were common in patients with methylmalonic acidaemia. It is likely that they are linked and essentially secondary to the tubulo-interstitial nephritis that is histologically demonstrable on renal biopsy. The failure of urinary concentrating ability and the disturbed urine acidification will contribute to the metabolic derangement during episodes of decompensation.


Plasmodium falciparum Malaria Parasites in Ghana Show Signatures of Balancing Selection at Artemisinin Resistance Predisposing Background Genes.
Sub-Saharan Africa is courting the risk of artemisinin resistance (ARTr) emerging in Plasmodium falciparum malaria parasites. Current molecular surveillance efforts for ARTr have been built on the utility of P. falciparum kelch13 (pfk13) validated molecular markers. However, whether these molecular markers will serve the purpose of early detection of artemisinin-resistant parasites in Ghana is hinged on a pfk13 dependent evolution. Here, we tested the hypothesis that the background pfk13 genome may be present before the pfk13 ARTr-conferring variant(s) is selected and that signatures of balancing selection on these genomic loci may serve as an early warning signal of ARTr. We analyzed 12 198 single nucleotide polymorphisms (SNPs) in Ghanaian clinical isolates in the Pf3K MalariaGEN dataset that passed a stringent filtering regimen. We identified signatures of balancing selection in 2 genes (phosphatidylinositol 4-kinase and chloroquine resistance transporter) previously reported as background loci for ARTr. These genes showed statistically significant and high positive values for Tajima's D, Fu and Li's F, and Fu and Li's D. This indicates that the biodiversity required to establish a pfk13 background genome may have been primed in clinical isolates of P. falciparum from Ghana as of 2010. Despite the absence of ARTr in Ghana to date, our finding supports the current use of pfk13 for molecular surveillance of ARTr in Ghana and highlights the potential utility of monitoring malaria parasite populations for balancing selection in ARTr precursor background genes as early warning molecular signatures for the emergence of ARTr.


Development of a Questionnaire to Assess Knowledge and Perceptions about Edible Insects.
Edible insects (EI) have been consumed as traditional foods in many parts of the globe, but in other regions, they are not readily accepted, particularly in Western countries. However, because EI are suggested to constitute a more sustainable protein food as compared with other sources of animal protein, they can be considered a future food that could help mitigate hunger and malnutrition. Additionally, new gastronomic trends are already targeting this area for exploring new potentialities. The objective of this work was to develop and validate a questionnaire to assess consumers' perceptions and knowledge about EI in seven different domains: D1. Culture and Tradition, D2. Gastronomic Innovation and Gourmet Kitchen, D3. Environment and Sustainability, D4. Economic and Social Aspects, D5. Commercialization and Marketing, D6. Nutritional Aspects and D7. Health Effects. The 64 items were subjected to item analysis and reliability analysis for validation, and factor analysis was also conducted to identify a grouping structure. The results validated all the items of the seven subscales with high values of Cronbach's alpha (α = 0.732 for D1, α = 0.795 for D2, α = 0.882 for D3, α = 0.742 for D4, α = 0.675 for D5, α = 0.799 for D6 and α = 0.788 for D7). However, by eliminating 17 items, the final values of the alpha increased in all subscales. Factor analysis with extraction by principal component analysis with varimax rotation extracted 14 factors that explained, in total, 65% of the variance, although the first two factors were the most important (35.7% variance explained). In conclusion, the confirmed usefulness of the questionnaire has been hereby validated for assessing consumer perceptions of and knowledge about EI.


Cryoprotective dehydration is widespread in Arctic springtails.
Cryoprotective dehydration (CPD) is a cold tolerance strategy employed by small invertebrates that readily lose water by evaporation when subjected to sub-zero temperatures in the presence of ice. Until now, relatively few species have been investigated using methods by which CPD can be shown. In the present study we investigated the cold tolerance strategy of seven soil arthropod species from the high Arctic Spitzbergen, and compared water content and water loss, body fluid melting points (MP) and survival under cold and desiccating conditions. We tested the hypothesis that CPD is a commonly occurring cold hardiness strategy among soil arthropods. We found that four springtail species (Hypogastrura viatica, Folsomia quadrioculata, Oligaphorura groenlandica and Megaphorura arctica; Collembola) went through severe dehydration and MP equilibration with ambient temperature, and thus overwinter by employing CPD, whereas a beetle (Atheta graminicola) and one of the springtails (Isotoma anglicana) were typical freeze avoiding species over-wintering by supercooling. Desiccation tolerance of the red velvet mite (Neomolgus littoralis) was also investigated; very low water loss rates of this species indicated that it does not survive winter by use of CPD. All in all, the results of the present study confirm the hypothesis that CPD is an effective over-wintering strategy which is widespread within soil arthropods.


Molecular characterization of an Akabane virus isolate from West Java, Indonesia.
We isolated an arbovirus from bovine blood in Indonesia. The arbovirus was obtained from the plasma of a cow showing no clinical symptoms in West Java in February 2014, and was identified as Akabane virus (AKAV) by AKAV-specific RT-PCR and subsequent sequence analysis. Phylogenetic analysis based on partial S segment indicated the AKAV isolate, WJ-1SA/P/2014, was most closely related with two isolates from Israel and Turkey reported in 2001 and 2015, respectively, and that WJ-1SA/P/2014 isolate belongs to AKAV genogroup Ib. This is the first isolation of AKAV from Indonesia.


Transcatheter Aortic Valve Implantation: Upcoming New Devices.
Transcatheter aortic valve implantation (TAVI) is an alternative to surgical aortic valve replacement in high-risk patients with symptomatic severe aortic stenosis. Several upcoming devices will optimize and facilitate the procedure: periprosthetic aortic regurgitation will be reduced by larger prosthesis sizes and repositionable next-generation transcatheter heart valves. Alternative access routes and smaller sheaths will reduce the rate of vascular complications. Cerebral protection devices might reduce the rate of strokes and silent cerebral embolism. Abandonment of balloon valvuloplasty and rapid ventricular pacing might facilitate the procedure and improve outcome. Thus, younger and healthier patients could benefit from TAVI in the near future.


Association Between Spikes in External Training Load and Shoulder Injuries in Competitive Adolescent Tennis Players: The SMASH Cohort Study.
Few studies have examined the association between the acute:chronic workload ratio (ACWR) and complaints/injuries in young tennis players. Primary aims of this study were to investigate if accumulated external workload "spikes" in ACWR of tennis training, match play, and fitness training, and to see if high or low workload/age ratio were associated with the rate of shoulder complaints/injuries in competitive adolescent tennis players. Additional aims were to report the incidence of complaints/injuries stratified by sex and level of play and to describe shoulder injury characteristics. Rapid increases in external workload are associated with the incidence of shoulder complaints and injuries. A cohort study. Level 3. At baseline, 301 adolescent competitive tennis players, 13 to 19 years, were screened and followed weekly for 52 weeks with questionnaires, in the years 2018 to 2019. Information about time-varying accumulated external workload spikes (uncoupled ACWR >1.3), and workload/age ratio, in 252 uninjured players were used in Cox regression analyses with the outcomes shoulder complaints (≥20) and injuries (≥40) (Oslo Sports Trauma Research Center Overuse Injury Questionnaire). For each additional workload spike in tennis training/match play, the hazard rate ratio (HRR) was 1.26 (95% CI, 1.13-1.40) for a shoulder complaint and 1.26 (95% CI, 1.15-1.39) for a shoulder injury. The HRR for fitness training was 1.11 (95% CI, 1.02-1.20) for a shoulder complaint and 1.18 (95% CI, 1.09-1.27) for a shoulder injury. Workload/age ratio was not associated with the rate of shoulder complaints or injuries. Accumulated external workload spikes of tennis training, match play, and/or fitness training are associated with a higher rate of shoulder complaints and shoulder injuries in competitive adolescent tennis players. Consistency in training load on a weekly basis is most likely more beneficial for adolescent tennis players regarding shoulder complaints/injuries than a training schedule comprising rapid increases (ie, spikes) in workload.


On the developmental properties and tissue interactions of hexokinase.
The interactions of the isozymes of hexokinase with cellular structure have been studied in the major tissues of the mouse during development. Overall, these data provide a clear indication that interactions between hexokinase and cellular structure are appreciable in all major tissues and at all stages of development, and an analysis of the isozyme status of the enzyme in both soluble and bound compartments has been effected. Further evidence in support of the already well documented interaction of hexokinase I to subcellular material in adult brain and kidney tissues is provided and extended to show that such interactions are extensive in both these tissues throughout development. In addition, evidence is provided that considerable hexokinase II activity is present in mouse foetal tissues in both the soluble and bound fractions and this isozyme is also shown to be the predominant "bound" form of the enzyme in adult skeletal muscle. By contrast, hexokinase III and IV are shown to be largely located in the cytosolic fraction of liver. The metabolic implications of these enzyme-structural interactions during development are discussed, as is the possibility of a functional linkage between hexokinase, which is bound to the mitochondria, and other enzymic components of the glycolytic sequence.


Cost-effectiveness of video-observed therapy for ambulatory management of active tuberculosis during the COVID-19 pandemic in a high-income country.
The coronavirus 2019 (COVID-19) pandemic caused suspension of directly observed therapy (DOT) for patients with active tuberculosis (TB). This study aimed to estimate the outcomes of pandemic-related DOT suspension and the cost-effectiveness of video-observed therapy (VOT) during the pandemic. A decision-analytic model was constructed to project outcomes of adult patients with active TB from the perspective of a US healthcare provider. Two model-based analyses were conducted: (1) before (with DOT) and during [with self-administered therapy (SAT)] the pandemic; and (2) VOT vs SAT during the pandemic. The primary outcome measures were direct medical costs and disability-adjusted life years (DALYs). In the base-case analysis, care during the pandemic (with SAT) increased the cost (by US$285 per patient) and DALYs (by 0.2155 per patient) in comparison with DOT. Care with VOT reduced DALYs (by 0.4870) and costs (by US$1797) in comparison with SAT. On probabilistic sensitivity analysis, care during the pandemic (with SAT) increased DALYs in 100% of 10,000 simulations, and increased costs in 55.52% of instances. Care with VOT reduced DALYs and costs in 99.7% and 68.79% of instances, respectively. The probability of VOT being cost-effective was 99.4% at the willingness-to-pay threshold of 50,000 US$/DALY. Suspension of DOT during the COVID-19 pandemic worsened treatment outcomes. VOT was found to be a cost-effective option for active TB care in an outpatient setting.


Divergent relationship between brain structure and cognitive functioning in patients with prominent negative symptomatology.
Investigating commonalities in underlying pathology of cognitive dysfunction and negative symptoms in schizophrenia is important, as both are core features of the disorder and linked to brain structure abnormalities. We aimed to explore the relationship between cognition, negative symptoms and brain structure in schizophrenia. A total of 225 patients with Schizophrenia spectrum disorder and 283 healthy controls from the Norwegian Thematically Organized Psychosis (TOP) cohort were included in this study. Patients were categorized into four patient subgroups based on severity of negative symptoms: no-negative- (NNS), threshold-negative- (TNS), moderate-negative- (MNS), and prominent-negative (PNS) subgroups. MRI measures of brain volume, mean cortical thickness and surface area from pre-selected brain regions were tested for relationships with general cognitive ability and negative symptom subgroups. Positive associations were found between brain volume, thickness, surface area and cognition in the dorsolateral prefrontal cortex (DLPFC), orbitofrontal cortex (OFC), fusiform gyrus (FG) and the left anterior cingulate cortex (ACC), but with no differences between subgroups. In the PNS subgroup, cognition was conversely negatively associated with brain volume in the left ACC. These results indicate that patients with prominent negative symptoms have different associations between cognition and brain structure in the left ACC, which may point to abnormal neurodevelopment.


Frontal electromyographic feedback. Stress attenuation and generalization.
This study evaluated the effects of one session of frontal electromyographic (EMG) feedback on (1) frontal EMG, (2) frontal EMG response to stress, (3) cardiovascular variables, and (4) cardiovascular responses to stress. Eighteen male and female undergraduate volunteers received either frontal EMG feedback or a relaxation instructions control procedure and were then exposed to a fear stimulus (visualization of a feared situation) and a post-stress adaptation period while several cardiovascular measures were monitored. In comparison to the control group, frontal EMG feedback significantly reduced resting levels of frontal EMG and frontal EMG response to stress but had no significant effect on cardiovascular measures. The results of this study suggest that one session of frontal EMG feedback may attenuate response to stress but, within the paradigm utilized, may be confined to the specific muscle groups monitored. Additional areas of needed research were noted including individual differences in generalization, the effects of EMG feedback from multiple sites sequentially and concomitantly, and the generalized effects from symptom-specific sites.


100 years of iodine testing of the cervix: A critical review and implications for the future.
We aim to describe the history of iodine testing of the cervix and identify areas where further work is required. We conducted a search of PubMed and Google Scholar. Full article texts were reviewed. Reference lists were screened for additional articles and books. 37 basic articles in journals including ones written in German and three basic articles in books were identified. Glycogen staining of the ectocervical squamous epithelium with iodine goes back to Paul Ehrlich (1854-1915). Walter Schiller (1887-1960) examined nearly 200 different dyes and found that vital staining of the cervical squamous epithelium was best achieved with Lugol's iodine solution, which was indicated by Jean Guillaume Lugol (1786-1851) for disinfection of the vagina. In 1928 W. Lahm observed that the glycogen content of a squamous epithelium cell decreases as anaplasia increases. From the outset, H. Hinselmann included the iodine test in the minimum requirements for colposcopy. In 1946 H. J. Wespi first mentioned the finding of an "uncharacteristic iodine negative area." The first international colposcopic terminology from Graz in 1975 lists the "iodine light area" among the different colposcopy findings. The IFCPC nomenclatures from Rome 1990, Barcelona 2002, and Rio de Janeiro 2011 have evaluated the iodine test and classified their findings differently. A breakthrough to effective cervical cancer screening in resource-limited settings in Africa, India, and Latin America was achieved with R. Sankaranarayanan's publication on naked-eye visual inspection of the cervix after application of Lugol's iodine. This paper is a step toward a better understanding of what we think and do today with iodine testing and what problems and upcoming tasks will arise in future.


Mediating Role of Labor on the Relationship Between Prenatal Psychopathologic Symptoms and Symptoms of Postpartum Depression in Women Who Give Birth Vaginally.
To investigate how prenatal symptoms of anxiety and depression and clinical aspects of labor (duration of labor, administration of oxytocin and epidural analgesia) interact with each other to contribute to symptoms of postpartum depression in women who give birth vaginally. A longitudinal design with measurement at three different time points: Time 1, 31 to 32 weeks gestation; Time 2, the day of labor and birth; and Time 3, 1 month after birth. Maternity ward of the Misericordia e Dolce Hospital in Prato, Italy. A total of 186 women at 31 to 32 weeks gestation were recruited during childbirth preparation courses at the maternity ward. At Time 1, women completed the Beck Depression Inventory and the State portion of the State-Trait Anxiety Inventory. At Time 2, midwives recorded clinical data related to labor, including duration of labor and administration of oxytocin and epidural analgesia. At Time 3, the women completed the Edinburgh Postnatal Depression Scale. Structural equation modeling was performed. Symptoms of depression (β = 0.36; p < .001; 95% confidence interval [CI] [0.17, 0.49]) and state anxiety (β = 0.25; p < .001; 95% CI = [0.04, 0.27]) during pregnancy positively affected symptoms of depression after birth. Greater levels of these prenatal symptoms predicted a more complicated labor (depression: β = 0.29; p < .01; 95% CI [0.00, 0.19]; anxiety: β = 0.30; p < .01; 95% CI [0.01, 0.14]), which, in turn, positively predicted greater levels of symptoms of depression at 1 month after birth (β = 0.34; p < .001; 95% CI [0.38, 1.51]). Moreover, results highlighted indirect effects that high levels of anxiety symptoms during pregnancy have on postpartum symptoms of depression through the clinical aspects of labor (β = 0.10; p < .01; 95% CI [0.00, 0.13]). These indirect effects were not significant for antenatal symptoms of depression (β = 0.10; p < .05; 95% CI [-0.02, 0.20]). Our findings confirm that symptoms of anxiety and depression during pregnancy represent significant risk factors for the clinical aspects of labor and for the development of symptoms of postpartum depression in the first month after childbirth.


[Survival Analysis of 76 Cases of Head and Neck Adenoid Cystic Carcinoma with Lung Metastasis].
Adenoid cystic carcinoma (ACC) of the head and neck often develops lung metastasis. At present, there are not many research reports on ACC lung metastasis, little is known about its exact clinical features and treatment results, and there is no consensus on the best treatment strategy. This study explored the effective treatment strategies, clinical outcomes and long-term prognosis of head and neck ACC lung metastases. The clinical and follow-up data of 76 patients with head and neck ACC lung metastases were retrospectively analyzed. According to the initial treatment of patients, they are divided into 4 groups: surgery, surgery+chemotherapy or radiotherapy, chemotherapy or radiotherapy and supportive treatment. The patients were staged according to the International Registry of Lung Metastases Staging System (IRLM). Kaplan-Meier method and Log-rank test were used to compare the statistical differences of overall survival (OS) and progression-free survival (PFS) of patients with different treatment methods and different IRLM stages. The OS and PFS of patients undergoing surgery are better than those of supportive therapy or radiotherapy and/or chemotherapy (OS: P<0.000,1; PFS: P<0.000,1). The OS and PFS of patients with low stage IRLM are better than those with high stage (OS: P<0.000,1; PFS: P<0.000,1). Patients with single lung metastasis and without pleural effusion have better OS and PFS. The long-term prognosis of patients with lung metastasis of head and neck ACC who undergo surgery is better than other treatments, which is related to higher OS and PFS. For patients with ACC lung metastases who are operationally eligible, the significance of complete surgical resection should be higher than other treatment options.


Amorphization-induced surface electronic states modulation of cobaltous oxide nanosheets for lithium-sulfur batteries.
Lithium-sulfur batteries show great potential to achieve high-energy-density storage, but their long-term stability is still limited due to the shuttle effect caused by the dissolution of polysulfides into electrolyte. Herein, we report a strategy of significantly improving the polysulfides adsorption capability of cobaltous oxide by amorphization-induced surface electronic states modulation. The amorphous cobaltous oxide nanosheets as the cathode additives for lithium-sulfur batteries demonstrates the rate capability and cycling stability with an initial capacity of 1248.2 mAh g-1 at 1 C and a substantial capacity retention of 1037.3 mAh g-1 after 500 cycles. X-ray absorption spectroscopy analysis reveal that the coordination structures and symmetry of ligand field around Co atoms of cobaltous oxide nanosheets are notably changed after amorphization. Moreover, DFT studies further indicate that amorphization-induced re-distribution of d orbital makes more electrons occupy high energy level, thereby resulting in a high binding energy with polysulfides for favorable adsorption.


Optimal Levels of Meloidogyne incognita Inoculum for Infection of Tomato and Peach in Vitro.
Penetration of second-stage juveniles (J2) of Meloidogyne incognita into tomato root explants and in vitro propagated peach plantlet roots were compared. Five inoculum levels were used: 25, 50, 75, 100, and 200 J2 for tomato; and 50, 100, 200, 500, and 1,000J2 for peach. The greatest root penetration into tomato was 30% at the 75 J2 level, but the maximum penetration into peach roots was only 8% at the 200 J2 level. The difference (P = 0.05) in penetration of M. incognita at all inoculum levels into these two hosts indicates that penetration versus inoculum density for in vitro studies need to be determined for different plant species.


It is all about the insects: a retrospective on 20 years of forensic entomology highlights the importance of insects in legal investigations.
This study highlights the importance of insect evidence by evaluating 949 insect-associated cases, including 139 entomological reports, from 2001 to 2019 at the Institute of Legal Medicine Frankfurt/Germany. With a high number of cases in the summer months and a low number in the colder season, 78.5% of the bodies were found indoors, regardless of year or month. In more than 80% of the cases, where PMI information was available (n = 704), the presumed PMI ranged from 1 to 21 days, a period during which entomological evidence can provide a day-specific estimate of PMImin. In cases where insects have been identified to species level (n = 279), most bodies were infested by one or two species with a maximum of 10 different species. Overall, a total of 55 insect species were found. Information on biology, activity and distribution of the most abundant taxa is given and applied for 5 case histories estimating different PMImins of up to over 6 months. Despite proved importance and scientific development of forensic entomology, insects are still rarely considered as a tool in forensic case work. The main reasons are a lack of awareness and (too) late involvement of a forensic entomologist. Our work shows that forensic entomology is an independent discipline that requires specialist expertise.


(-)-Menthylamine derivatives as potent and selective antagonists of transient receptor potential melastatin type-8 (TRPM8) channels.
A series of twenty-two (-)-menthylamine derivatives was synthesized and tested on TRPM8, TRPV1, and TRPA1 channels. Five of the novel compounds, that is, 1d, 1f, 2b, 2c, and 2e behaved as potent TRPM8 antagonists with IC(50) values versus icilin and (-)-menthol between 20 nM and 0.7 microM, and were between 4- and approximately 150-fold selective versus TRPV1 and TRPA1 activation. Compound 1d also induced caspase 3/7 release in TRPM8-expressing LNCaP prostate carcinoma cells, but not in non-TRPM8 expressing DU-145 cells. Five other derivatives, that is, 1a, 1g, 1h, 2f, and 2h were slightly less potent than previous compounds but still relatively selective versus TRPV1 and TRPA1.


Improved detection of donor-specific HLA-class II antibody in kidney transplant recipients by modified immunocomplex capture fluorescence analysis.
Immunocomplex capture fluorescence analysis (ICFA) which basic principle is same as Luminex crossmatch (LXM), could detect donor-specific HLA antibody (DSA). The advantages of ICFA are (i) detection of DSA and (ii) no requirement of viable cells over the flow cytometry crossmatch (FCXM). However, FCXM has been widely used because of its higher sensitivity than ICFA, in particular HLA-class II antibody detection. In this study the accuracy of DSA detection against HLA-class II was investigated by modifying the original method of ICFA. Increment of the sensitivity was found when purified peripheral blood mononuclear cells (PBMCs) were used instead of whole blood. An ICFA-PBMC in addition to FCXM-T/B was conducted for 118 patients before kidney transplantation and 13 patients with de novo DSA against HLA-class II after transplantation. Significantly positive correlation was observed between the values of ICFA-PBMC and DSA mean fluorescence intensity (MFI) targeting class II (p < 0.0001). When the cutoff level of 1.4 was determined by receiver operating characteristic curve analysis, the average DSA MFI was found to be significantly higher in the ICFA-PBMC (class II) positive group comparing to that in the negative group (12,217 vs 3885, p = 0.0027). ICFA-PBMC and optimized cutoff level could provide valid information in cases of suspected DSA.


Increased Posterior Tibial Slope and Meniscal Slope Could Be Risk Factors for Meniscal Injuries: A Systematic Review.
The purpose of this systematic review was to summarize the available evidence and examine the relation between the posterior tibial slope (PTS) and meniscal slope (MS) and the incidence of meniscal injury. PubMed, Embase, the Cochrane Central Register of Controlled Trials, and Web of Science were searched from inception to February 23, 2021. Cohort studies investigating the association between PTS or MS and the risk of meniscal injury were included. Two authors independently conducted the literature search, data extraction, and quality assessment. Sixteen studies with a total of 2,670 patients were included. For meniscal injury with an anterior cruciate ligament tear, the lateral PTS in the lateral meniscal root tear group (range, 8.0°-12.6°) was significantly higher than that in the control group (range, 4.0°-10.7°). Furthermore, there appeared to be a relation between a greater medial MS and the presence of a ramp lesion (range, 2.6°-6.7° for ramp lesion vs 2.0°-5.1° for control). For degenerative meniscal injury, the medial PTS in the medial meniscal posterior root tear group (range, 6.15°-10.4°) was significantly greater than that in the control group (range, 4.0°-9.8°). On the basis of the available evidence, for meniscal injury with an anterior cruciate ligament tear, an increased lateral PTS was associated with a higher risk of lateral meniscal tears and lateral meniscal posterior root tears. Furthermore, there appeared to be a relation between an increased medial MS and a higher risk of ramp lesions. For degenerative meniscal injury, most of the included studies showed that a larger medial PTS could increase the risk of medial meniscal tears and medial meniscal posterior root tears. Level III, systematic review of Level III studies.


Accessory stimulation in the time course of visuomotor information processing: stimulus intensity effects on reaction time and response force.
A series of three visual choice-reaction time experiments were performed to systematically investigate the effects of accessory auditory stimulation on response time (RT) and response force (RF). In Experiment 1, the effect of accessory auditory stimulation on early visual information processing was investigated. Experiments 2 and 3 were designed to examine the effects of accessory intensity on RT and RF across the entire time course of sensorimotor processing. Accessory stimulation accelerated response speed only when presented within 100 ms after onset of the visual response signal. An enhancing effect of accessory stimulation on RF, however, was found as late as 220 ms after onset of the response signal. These findings support the notion that response speed and response dynamics represent functionally independent sensorimotor phenomena.


Long-term safety of pembrolizumab monotherapy and relationship with clinical outcome: A landmark analysis in patients with advanced melanoma.
Long-term safety of pembrolizumab in melanoma was analyzed in KEYNOTE-001, KEYNOTE-002, and KEYNOTE-006. Analysis involved patients who received ≥1 pembrolizumab dose. Lead-time bias was addressed via landmark analyses in patients who were progression-free before day 147. Adverse events (AEs) were analyzed for 1567 patients (median follow-up, 42.4 months). Most AEs were mild/moderate; grade 3/4 treatment-related AEs occurred in 17.7% of patients. Two pembrolizumab-related deaths occurred. Any-grade immune-mediated AEs (imAEs) occurred in 23.0%, most commonly hypothyroidism (9.1%), pneumonitis (3.3%), and hyperthyroidism (3.0%); grade 3/4 imAEs occurred in 6.9% of patients. Most imAEs occurred within 16 weeks of treatment. In landmark analysis, patients who did (n = 79) versus did not (n = 384) develop imAEs had similar objective response rates (ORRs) (64.6% versus 63.0%); median time to response (TTR), 5.6 months for both; median duration of response (DOR), 20.0 versus 25.3 months; median progression-free survival (PFS), 17.0 versus 17.7 months; median overall survival (OS), not reached (NR) versus 43 months (p = 0.1104). Patients who did (n = 17) versus did not (n = 62) receive systemic corticosteroids had similar ORRs (70.6% vs. 62.9%) and median TTR (6.4 vs. 5.6 months) but numerically shorter median PFS (9.9 vs. 17.0 months); median DOR, 14.2 months versus NR; median OS, NR for both. These results enhance the knowledge base for pembrolizumab in advanced melanoma, with no new toxicity signals after lengthy follow-up of a large population. In landmark analyses, pembrolizumab efficacy was similar regardless of imAEs or systemic corticosteroid use. NCT01295827, NCT01704287, NCT01866319.


Human pharmacokinetics of the antiviral drug DHPG.
The pharmacokinetics of the antiviral drug 9-[2-hydroxy-1-(hydroxymethyl) ethoxymethyl]guanine (DHPG) were examined in six patients receiving 2.5 or 5.0 mg/kg every 8 or 12 hours for human cytomegalovirus (HCMV) pneumonitis or retinitis. Biexponential decay with a mean distribution t1/2 of 0.23 hours and terminal t1/2 of 2.53 hours was observed. Total clearance correlated well with and exceeded creatinine clearance by a factor of 2.4. Mean volume of the central compartment was 15.26 L/1.73 m2 and the volume of distribution at steady state was 32.8 L/1.73 m2. Peak (model predicted) and trough plasma concentrations were 4.75 to 6.20 micrograms/ml and less than 0.25 to 0.63 microgram/ml, respectively, in patients receiving 2.5 mg/kg. Peak concentrations are well above those needed to inhibit HCMV at the 50% level (ID50) and troughs are near this ID50. Cerebrospinal fluid concentrations of DHPG indicate a penetration of 24% to 67%. No accumulation of DHPG was apparent in these patients. However, dosage reduction is necessary in renal insufficiency. Neutropenia occurred in one patient. The plasma concentration profile of DHPG suggests potential beneficial activity against HCMV.


Individuals with self-determined motivation for eating have better overall diet quality: Results from the PREDISE study.
The study aimed at documenting motivational orientations for the regulation of eating as defined by self-determination theory and their association with sociodemographic characteristics and overall diet quality. As part of the PREDISE study, French-speaking women (n = 550) and men (n = 547), aged 18-65 years, living in the Province of Québec, Canada, completed online validated questionnaires. The Regulation of Eating Behavior Scale, based on the self-determination theory, assessed self-determined and non-self-determined motivation to regulate one's eating behavior. Three web-based 24-h food recalls were completed and used to compute the Canadian Healthy Eating Index 2007 (C-HEI), an indicator of the overall adherence to Canadian guidelines for healthy eating. Multiple linear regressions were performed to assess how regulation styles are associated with the C-HEI. Model 1 included no covariate, model 2 included sociodemographic covariates, and fully adjusted model 3 included as covariates sociodemographic variables as well as variables that were previously associated with diet quality, namely nutrition knowledge and social support for healthy eating. Women (p < 0.0001), older individuals (p = 0.0002), those with a higher education level (p < 0.0001), and non-smokers (p < 0.0001) reported higher self-determined motivation score than their counterparts. Self-determined motivation was positively (model 1: B = 4.67, p < 0.0001; model 2: B = 3.82, p < 0.0001; model 3: B = 3.61, p < 0.0001) and non-self-determined motivation was negatively (model 1: B = -1.62, p = 0.0009; model 2: B = -1.63, p = 0.0006; model 2: B = -1.49, p = 0.0022) associated with C-HEI. The present study suggests that some subgroups of the general adult population show more self-determined motivation for eating, which is associated with a better diet quality independently of individual characteristics and other individual and social determinants of healthy eating. Strategies to help individuals internalize the regulation of eating should be further investigated.


Safety and Feasibility of Immunoadsorption with Heparin Anticoagulation in Preparation of ABO-Incompatible Kidney Transplantation: A Retrospective Single-Center Study.
Immunoadsorption (IA) of isohemagglutinins is an often-crucial procedure in preparation of major ABO blood group-incompatible living donor kidney transplantation (ABOi LDKT). Standard citrate-based anticoagulation during the procedure has potential disadvantages for distinct patient groups. In this study, we report our experience with an alternative anticoagulation scheme using heparin during IA for selected patients. We conducted a retrospective analysis of all patients who underwent IA with heparin anticoagulation between February 2013 and December 2019 at our institution with focus on the safety and efficacy of the adapted procedure. For further validation, we compared graft function, graft survival, and overall survival with those of all recipients of living donor kidney transplants with or without pretransplant desensitizing apheresis for ABO antibodies at our institution during the same period. In thirteen consecutive patients prepared for ABOi LDKT with IA with heparin anticoagulation, no major bleeding or other significant complications were observed. All patients achieved sufficient isohemagglutinin titer reduction to proceed to transplant surgery. Graft function, graft survival, and overall survival did not significantly differ from patients treated with standard anticoagulation for IA or ABO compatible recipients of living donor kidneys. IA with heparin in preparation of ABOi LDKT is safe and feasible for selected patients after internal validation.


Th1 cytokine gene polymorphism and the corresponding plasma cytokine levels: A comparative study in HIV-1 positive and exposed uninfected infants.
The pro-inflammatory (Th1) cytokines namely interleukin (IL)-2, IL-6, IL-12, interferon (IFN)-γ, tumor necrosis factor-α (TNF-α) are vital in the clearance of HIV infection. This prospective cohort study aimed to evaluate the polymorphisms of Th1 cytokine genes and their corresponding plasma cytokine levels in HIV-1 positive and exposed uninfected (EU) infants born to HIV-1 positive mothers. CD4 count, viral load of HIV-1 positive mothers was done using commercially available reagents. Cytokine genotyping analysis and levels were done in 20 HIV-1 positive and 54 EU infants. The polymorphisms of Th1 cytokines were done using the PCR-SSP method. Plasma cytokine levels were estimated using Bio-Plex-Pro cytokine assay (BIO-RAD; USA). Results revealed treatment status of the mothers and viral load were the two confounding factors having a significant effect on HIV status of the infant. TNF-α GG genotype is significantly higher in EU infants as compared with HIV-1 positive infants. GG genotype was associated with high TNF- α levels in HIV-1 positive infants but the difference was not statistically significant. HIV-1 positive infants with -IFN-γ (+874) TT genotype was significantly associated with high IFN-γ levels. To the best of our knowledge, this is the first study reporting the role of Th1 cytokine gene polymorphisms and their corresponding plasma cytokine levels in HIV-1 positive and EU infants from India.


Automated classification of urine biomarkers to diagnose pancreatic cancer using 1-D convolutional neural networks.
Early diagnosis of Pancreatic Ductal Adenocarcinoma (PDAC) is the main key to surviving cancer patients. Urine proteomic biomarkers which are creatinine, LYVE1, REG1B, and TFF1 present a promising non-invasive and inexpensive diagnostic method of the PDAC. Recent utilization of both microfluidics technology and artificial intelligence techniques enables accurate detection and analysis of these biomarkers. This paper proposes a new deep-learning model to identify urine biomarkers for the automated diagnosis of pancreatic cancers. The proposed model is composed of one-dimensional convolutional neural networks (1D-CNNs) and long short-term memory (LSTM). It can categorize patients into healthy pancreas, benign hepatobiliary disease, and PDAC cases automatically. Experiments and evaluations have been successfully done on a public dataset of 590 urine samples of three classes, which are 183 healthy pancreas samples, 208 benign hepatobiliary disease samples, and 199 PDAC samples. The results demonstrated that our proposed 1-D CNN + LSTM model achieved the best accuracy score of 97% and the area under curve (AUC) of 98% versus the state-of-the-art models to diagnose pancreatic cancers using urine biomarkers. A new efficient 1D CNN-LSTM model has been successfully developed for early PDAC diagnosis using four proteomic urine biomarkers of creatinine, LYVE1, REG1B, and TFF1. This developed model showed superior performance on other machine learning classifiers in previous studies. The main prospect of this study is the laboratory realization of our proposed deep classifier on urinary biomarker panels for assisting diagnostic procedures of pancreatic cancer patients.