Changes in physical activity in pre-schoolers and first-grade children: longitudinal study in the Czech Republic.
The transition from kindergarten to first year at school is associated with a variety of psychosocial changes in children. The aim of this longitudinal study was to identify the changes in children's physical activity (PA) upon entry to first year at school; and to identify the days of the school week that exhibit low PA. We monitored the PA levels of 176 children twice: initially in kindergarten and again in first-year classes at school. The age (mean +/- standard deviation) of children at kindergarten was 5.7 +/- 0.5 years and 6.7 +/- 0.5 years at the first year of elementary school. We evaluated PA employing the activity energy expenditure (AEE - kcal/kg/day) from Caltrac accelerometer and daily amount of steps (STEPS) from Yamax pedometer. Participants were monitored over 7 days to include a weekend. The mean AEE was 11.5 in pre-school girls and 12.9 kcal/kg/day in boys; and STEPS were 9923 steps/day in girls and 11 864 in boys on weekdays. At weekends, it was 11.5 kcal/kg/day in girls and 12.7 kcal/kg/day in boys and 10 606 steps/day in girls and 11 182 steps/day in boys. The mean AEE and STEPS in first-grade girls and boys was 9.9 and 9.8 kcal/kg/day respectively, and 7911 and 8252 steps/day respectively on weekdays, and 8.8 and 9.0 kcal/kg/day and 6872 and 7194 steps/day respectively at weekends. First-grade school children had significantly lower PA than pre-school children on weekdays (P < 0.0001) and at weekends (P < 0.0001). Decline in PA on weekdays was during time spent at school (P < 0.0001) and not during after-school children's leisure time. The parts of the week when first-grade school children show low PA are the times spent in lessons and in after-school nursery and at weekends. PA needs to be promoted using intervention programmes mainly during the after-school nursery programmes and at weekends.


Natural Flavonoid Pectolinarigenin Alleviated Hyperuricemic Nephropathy via Suppressing TGFβ/SMAD3 and JAK2/STAT3 Signaling Pathways.
Natural flavonoid pectolinarigenin (PEC) was reported to alleviate tubulointerstitial fibrosis of unilateral ureteral obstruction (UUO) mice in our previous study. To further investigate nephroprotective effects of PEC in hyperuricemic nephropathy (HN), adenine and potassium oxonate induced HN mice and uric acid-treated mouse kidney epithelial (TCMK-1) cells were employed in the study. As a result, PEC significantly lowered serum uric acid level and restored hyperuricemia-related kidney injury in HN mice. Meanwhile, PEC alleviated inflammation, fibrosis, and reduced adipokine FABP4 content in the kidneys of HN mice and uric acid-treated TCMK-1 cells. Mechanistically, PEC inhibited the TGF-β1 expression as well as the phosphorylation of transcription factor SMAD3 and STAT3 to regulate the corresponding inflammatory and fibrotic gene expression in kidney tissues. In conclusion, our results suggested that PEC could inhibit the activation of SMAD3 and STAT3 signaling to suppress inflammation and fibrosis, and thereby alleviate HN in mice.


The diagnostic yield and complications of open lung biopsies in kidney transplant patients with pulmonary disease.
The purpose of this study was to assess the efficacy of open lung biopsy (OLB) in determining the specific diagnosis and the related complications in patients with undiagnosed diffuse pulmonary infiltrates. This single center, retrospective study included adult kidney transplant patients who underwent OLB. The patients had diffuse pulmonary infiltrates without definitive diagnoses and failed to respond to empiric antibiotic treatment. We analyzed the number of specific diagnoses, changes in treatment and the occurrence of complications in these patients. A logistic regression was used to determine which variables were predictors of hospital mortality. From April 2010 to April 2014, 87 patients consecutively underwent OLB. A specific diagnosis was reached in 74 (85.1%) patients. In 46 patients (53%), their therapeutic management was changed after the OLB results. Twenty-five (28.7%) patients had complications related to the OLB. The hospital mortality rate was 25.2%. Age, SAPS3 score and complications related to the procedure were independent predictors of all-cause mortality. OLB is a high-risk procedure with a high diagnostic yield in kidney transplant patients with diffuse pulmonary infiltrates who did not have a definitive diagnosis and who failed to respond to empiric antibiotic treatment. Complications related to OLB were common and were independently associated with intra-hospital mortality.


[Dysphagia due to giant cervical osteophytes].
We reported a patient with isolated dysphagia due to an esophageal canal stenosis compressed by focal cervical spondylotic osteophytes. The patient was a 63-year-old male who developed swallowing disturbance of predominantly solid materials. The neurological examination showed subjectively isolated dysphagia unassociated with any significant cranial nerve signs. Cervical magnetic resonance imaging and X-ray demonstrated giant cervical spurs focally at the ventral portion of the C 5/6 vertebral bodies, protruding ventrally and compressing mildly to the esophageal canal. The intervertebral disc formation was relatively preserved between the osteophytes. At the neck flexion portion, the esophagus was severely compressed between the osteophytes and the epiglottis. Videofluoroscopic examination of swallowing function showed a congestion of the contrast medium at the portion of the epiglottic vallecula, unassociated with aspiration into the trachea. The endoscopic examination showed erosion of the posterior wall of the esophagus at the portion compressed by the osteophytes. The anti-inflammatory medication improved the symptom of dysphagia, and the patient has been carefully followed-up without a surgical procedure. The presence of giant osteophyte should be considered in the differential diagnosis of subjectively isolated dysphagia unassociated with any significant cranial nerve signs.


Bacterial Extracellular Polysaccharides in Biofilm Formation and Function.
Microbes produce a biofilm matrix consisting of proteins, extracellular DNA, and polysaccharides that is integral in the formation of bacterial communities. Historical studies of polysaccharides revealed that their overproduction often alters the colony morphology and can be diagnostic in identifying certain species. The polysaccharide component of the matrix can provide many diverse benefits to the cells in the biofilm, including adhesion, protection, and structure. Aggregative polysaccharides act as molecular glue, allowing the bacterial cells to adhere to each other as well as surfaces. Adhesion facilitates the colonization of both biotic and abiotic surfaces by allowing the bacteria to resist physical stresses imposed by fluid movement that could separate the cells from a nutrient source. Polysaccharides can also provide protection from a wide range of stresses, such as desiccation, immune effectors, and predators such as phagocytic cells and amoebae. Finally, polysaccharides can provide structure to biofilms, allowing stratification of the bacterial community and establishing gradients of nutrients and waste products. This can be advantageous for the bacteria by establishing a heterogeneous population that is prepared to endure stresses created by the rapidly changing environments that many bacteria encounter. The diverse range of polysaccharide structures, properties, and roles highlight the importance of this matrix constituent to the successful adaptation of bacteria to nearly every niche. Here, we present an overview of the current knowledge regarding the diversity and benefits that polysaccharide production provides to bacterial communities within biofilms.


[Surgical treatment of presacral recurrent rectal cancer].
Presacral recurrent rectal cancer (PRRC) which involves the presacral fascia or/and sacral bone is one of the classification of locally recurrent rectal cancer. Presacral fascia or/and sacral bone involvement and posterior fixation make treatment difficult. In recent years, there are many researches on the diagnosis, surgical treatment and surgical methods of PRRC. The major therapeutic principle of PRRC is to perform radical resection with combined therapy under the evaluation of the multidisciplinary team (MDT). Among the surgical methods, abdominal resection or abdominoperineal resection, sacrectomy and abdominosacral resection are the common surgical methods for the treatment of PRRC. Recent articles revealed that limited sacrectomy could also result in good efficacy, provided strict indications for this procedure. With the development of minimally invasive technology, the application of laparoscopic technology in PRRC has been increasingly emphasized. In summary, the surgical principle of PRRC has shifted from solely pursuring radical resection to obtaining good efficacy as well as reduced morbidity with individualized management. In this article, the research progress of surgical methods of presacral recurrent rectal cancer in recent years are reviewed in order to provide reference to clinical diagnosis and treatment.


Stability in a changing world - palm community dynamics in the hyperdiverse western Amazon over 17 years.
Are the hyperdiverse local forests of the western Amazon undergoing changes linked to global and local drivers such as climate change, or successional dynamics? We analyzed local climatic records to assess potential climatic changes in Yasuní National Park, Ecuador, and compared two censuses (1995, 2012) of a palm community to assess changes in community structure and composition. Over 17 years, the structure and composition of this palm community remained remarkably stable. Soil humidity was significantly lower and canopy conditions were significantly more open in 2012 compared to 1995, but local climatic records showed that no significant changes in precipitation, temperature or river level have occurred during the last decade. Thus, we found no evidence of recent directional shifts in climate or the palm community in Yasuní. The absence of changes in local climate and plant community dynamics in Yasuní contrasts with recent findings from eastern Amazon, where environmental change is driving significant changes in ecosystem dynamics. Our findings suggest that until now, local forests in the northwest Amazon may have escaped pressure from climate change. The stability of this rich palm community embedded in the hyperdiverse Yasuní National Park underlines its uniqueness as a sanctuary for the protection of Amazonian diversity from global change impacts.


Persistent loneliness due to COVID-19 over 18 months of the pandemic: A prospective cohort study.
Loneliness was common early in the COVID-19 pandemic due to physical distancing measures, but little is known about how loneliness persisted into later stages of the pandemic. We therefore examined longitudinal trajectories of loneliness over 18 months of the pandemic and subgroups at risk for persistent loneliness. We used data from the COVID-19 & Chronic Conditions study collected between March 27, 2020 to December 10, 2021, including 641 predominantly older adults with ≥1 chronic condition who completed six interviews at approximately 3 month intervals. Participants reported loneliness (defined as some, most, or all of the time) during the past week due to COVID-19. We used trajectory mixture models to identify clusters of individuals following similar trajectories of loneliness, then determined subgroups likely to be classified in different loneliness trajectories using multivariable regression models adjusted for sociodemographic and clinical covariates. Participants were on average 63 years old, 61% female, 30% Black, 20% Latinx, and 29% were living below the poverty level. There was an overall reduction in loneliness over time (March to April/2020: 51% to September to December/2021: 31%, p = 0.01). Four distinct trajectory groups emerged: (1) "Persistent Loneliness" (n = 101, 16%); (2) "Adapted" (n = 141, 22%), individuals who were initially lonely, with feelings of loneliness decreasing over time; (3) "Occasional loneliness" (n = 189, 29%); and (4) "Never lonely" (n = 211, 33%). Subgroups at highest risk of the "Persistently Lonely" trajectory included those identifying as Latinx (aOR 2.5, 95% CI: 1.2, 5.2), or living in poverty (aOR 2.5; 95% CI: 1.4, 4.6). Although loneliness declined for a majority of older adults during the pandemic in our sample, persistent loneliness attributed to the COVID-19 pandemic was common (1 in 6 adults), particularly among individuals identifying as Hispanic/Latinx or living in poverty. Interventions addressing loneliness can ease pandemic-related suffering, and may mitigate long-term mental and physical health consequences.


Ultrasound diagnosis of the Pena Shokeir phenotype at 14 weeks of pregnancy.
This report describes the early prenatal diagnosis of the Pena Shokeir phenotype in an at-risk patient at 14 weeks' gestation. The diagnosis was based on an abnormal fetal movement profile, in association with an abnormal position of the fetal limbs. Pena Shokeir phenotype describes an inherited condition characterized by arthrogryposis and dysmorphic features as a result of fetal akinesia. It is a lethal abnormality and early diagnosis allows safer surgical methods of termination.


Single-center management of 136 patients with cancer of unknown primary site (CUP syndrome) over a period of 10 years.
Cancer of unknown primary site (CUP syndrome) is a particular challenge in oncology which occurs in about 5-10% of cancer patients. Here, we investigated clinicopathological and prognostic factors in patients with CUP syndrome in a retrospective analysis. 136 patients with CUP syndrome who were treated in our Department of Oncology and Hematology were analyzed over a period of 10 years. Clinical and histopathological characteristics, response to chemotherapy, survival and prognostic factors were investigated in a retrospective analysis. 83 of the patients (61%) received first-line chemotherapy, which induced an overall response rate of 19%. Altogether 37 different chemotherapy regimens were used. Median overall survival of all patients was 7.9 months. In multivariate Cox regression analysis, gender, Karnofsky performance status, treatment modality and extent of disease were identified as independent prognostic factors. Our analysis showed a poor prognosis for patients with CUP syndrome. The response rate to chemotherapy was low with no significant benefit for any of the investigated cytotoxic agents. Newer diagnostic and therapeutical approaches might contribute to an improvement of prognosis, and their value is currently investigated in prospective studies.


NutritionDay 2010 audit in Jinling hospital of China.
NutritionDay is an annual worldwide cross-sectional multicentre audit. This report aimed to describe the results of nutritionDay 2010 in Jinling hospital, providing a map of the prevalence of malnutrition and actual nutrition therapy practice in different units. The risk factors to malnutrition and length of hospital stay were also investigated. The data was collected from 233 inpatients from Jinling hospital on Nov 4th, 2010, using standardized questionnaires. Malnutrition was objectively defined as BMI <20 or unintentional weight loss >5% in the past three months. Risk factors for malnutrition and the impact of multiple factors on length of hospital stay were analyzed. 233 inpatients participated in this audit (143 M: 90 F; mean±SD age 50.6±18.5 years). Of the patients, 42.5% were malnourished. Multivariable analysis revealed three risk factors for malnutrition: gender, food intake and length of hospital stay before audit. Longer length of hospital stay prevailed in patients aged >=65 years (p<0.001), and there was a positive and significant, but weak, correlation between length of hospital stay and age. The prevalence of malnutrition was high. Higher age may be the main contributor to longer length of hospital stay. This was the first study to obtain data from hospitalized patients' nutritional status in China during the nutritionDay audit and the valuable results could supply evidence for clinical nutrition support.


Dispersal depression with habitat fragmentation in the bog fritillary butterfly.
Habitat fragmentation is expected to impose strong selective pressures on dispersal rates. However, evolutionary responses of dispersal are not self-evident, since various selection pressures act in opposite directions. Here we disentangled the components of dispersal behavior in a metapopulation context using the Virtual Migration model, and we linked their variation to habitat fragmentation in the specialist butterfly Proclossiana eunomia. Our study provided a nearly unique opportunity to study how habitat fragmentation modifies dispersal at the landscape scale, as opposed to microlandscapes or simulation studies. Indeed, we studied the same species in four landscapes with various habitat fragmentation levels, in which large amounts of field data were collected and analyzed using similar methodologies. We showed the existence of quantitative variations in dispersal behavior correlated with increased fragmentation. Dispersal propensity from habitat patches (for a given patch size), and mortality during dispersal (for a given patch connectivity) were lower in more fragmented landscapes. We suggest that these were the consequences of two different evolutionary responses of dispersal behavior at the individual level: (1) when fragmentation increased, the reluctance of individuals to cross habitat patch boundaries also increased; (2) when individuals dispersed, they flew straighter in the matrix, which is the best strategy to improve dispersal success. Such evolutionary responses could generate complex nonlinear patterns of dispersal changes at the metapopulation level according to habitat fragmentation. Due to the small size and increased isolation of habitat patches in fragmented landscapes, overall emigration rate and mortality during dispersal remained high. As a consequence, successful dispersal at the metapopulation scale remained limited. Therefore, to what extent the selection of individuals with a lower dispersal propensity and a higher survival during dispersal is able to limit detrimental effects of habitat fragmentation on dispersal success is unknown, and any conclusion that metapopulations would compensate for them is flawed.


Student characteristics and subculture trends in interpersonal skills workshops.
The aim of this study was to explore the experiences of student nurses when learning interpersonal skills and to examine the impact of small group work. A qualitative methodology was used. The sample comprised 50 student nurses who were undertaking the preregistration diploma course. In-depth interviews were employed to collect the data. The findings indicated that many students resisted learning owing to a range of personal characteristics that influenced the learning process. A student typology was constructed which highlighted four types of student characteristics that emerged from the data. Significant characteristics relating to each type of student behaviour are emphasized. This report also combines the student typology with the emergence of three distinct subcultures that frequently both interrupt and enhance learning. In presenting the subcultures the implications for group work are considered in conjunction with study limitations.


Production, characteristics and applications of the cell-bound phytase of Pichia anomala.
Among several yeasts isolated from dried flowers of Woodfordia fruticosa, Pichia anomala produced a high titre of cell-bound phytase. The optimization of fermentation variables led to formulation of media and selection of cultural variables that supported enhanced phytase production. The enzyme productivity was very high in fed batch fermentation in air-lift fermentor as compared to that in stirred tank fermentor. Amelioration in the cell-bound phytase activity was observed when yeast cells were permeabilized with Triton-X-100. The enzyme is thermostable and acid stable with broad substrate specificity, the characteristics that are desirable for enzymes to be used in the animal feed industry. The phytase-encoding gene was cloned and sequenced. The 3D structure of the enzyme was proposed by comparative modeling using phytase of Debaryomyces occidentalis (50% sequence identity) as template. When broiler chicks, and fresh water and marine fishes were fed with the feed supplemented with yeast biomass containing phytase, improvement in growth and phosphorus retention, and decrease in the excretion of phosphorus in the faeces were recorded. The cell-bound phytase of P. anomala could effectively dephytinize wheat flour and soymilk.


Cleavage of dT8 and dT8 phosphorothioyl analogues by Escherichia coli DNA topoisomerase I: product and rate analysis.
Escherichia coli DNA topoisomerase I catalyzes the cleavage of short, single-stranded oligodeoxynucleotides with dT8 as the shortest cleavable oligo(thymidylic acid). The 5'-32P-labeled products formed from the cleavage of [5'-32P]dT8 are dT5, dT4, and dT3 with over 70% of the substrate cleaved to dT4. Mg(II) ions affect this product distribution by increasing the percentage of dT4 formed. The substitution of a sulfur atom for a nonbridging oxygen atom in a phosphodiester linkage yields oligodeoxynucleotide phosphorothioyl (PS) analogues. The epimers of the analogues were separated, and the position and stereochemistry of the phosphorothiodiester bond were determined. Topoisomerase I is stereospecific in its reactivity toward these analogues. With the oligodeoxynucleotide PS analogue substrates, the rate of cleavage, the stereospecificity, and the product distribution depend upon the position and the stereochemistry of the phosphorothiodiester linkage.


Relation between retinal vein occlusions and axial length.
To evaluate the ocular axial length as a risk factor for development of central retinal vein occlusion (CRVO) and branch retinal vein occlusion (BRVO). Ocular axial lengths were measured, by A-scan ultrasonography, in 17 patients with CRVO and 41 patients with BRVO and compared with those of contralateral unaffected eyes and 66 age matched controls. In 17 patients with CRVO the mean axial length of affected eyes was 22.25 (SD 0.19) mm and of unaffected eyes was 22.61 (0.13) mm. In 41 patients with BRVO the mean axial length of affected eyes was 22.89 (0.11) mm and of unaffected eyes was 22.99 (0.12) mm. These findings confirm that the axial lengths in CRVO and BRVO were significantly shorter than in the controls. This significant difference may be a risk factor in the development of CRVO and BRVO.


Presence of iceA1 but not cagA, cagC, cagE, cagF, cagN, cagT, or orf13 genes of Helicobacter pylori is associated with more severe gastric inflammation in Taiwanese.
The presence of the cag pathogenicity island (PAI) may enhance the virulence of Helicobacter pylori. This study determined the prevalence of genotypes of the PAI, IS605, and iceA1 in H. pylori strains in Taiwanese and whether these genotypes were related to the type of clinical disease or gastric pathology. One hundred clinical isolates were collected from 33 duodenal ulcer, 41 nonulcer related dyspepsia, 14 gastric ulcer, and 12 gastric malignancy patients. Polymerase chain reaction (PCR) assays were performed for cagA, cagC, cagE, cagF, and cagN in the cagI region, cagT and orf13 in the cagII region, and IS605 and iceA1 in all H. pylori isolates. Gastric histology of the infected host was reviewed using the updated Sydney system. All strains were positive for all the selected genes in the PAI. PCR amplification found IS605 in 17%, while colony hybridization revealed it in 36% of strains. The prevalence of the cagA gene detected by PCR using cagA1, cagA2, and cagA3 primers was 26, 100, and 100%, respectively. The iceA1 gene existed in 72% of the H. pylori isolates. The mean ulcer size and the severity of acute gastric inflammation in patients infected with iceA1-positive strains were significantly greater than in those infected with iceA1-negative strains (p < 0.05). All clinical H. pylori isolates from different gastric diseases in our Taiwanese patients were positive for the PAI, but only 36% of these isolates carried an IS605 insertion. The selected genes in the PAI were not correlated with disease outcome. The determination of cagA prevalence is based on the selection of suitable primers. In contrast, bacterial factors such as the presence of iceAl could be related to severity of gastric inflammation and increase in ulcer size in H. pylori-infected patients.


Portacaval shunt control animals: physiological consequences derived from the sham operation.
The portacaval shunt in the rat is a frequently used experimental model of portosystemic encephalopathy. Among other consequences of this surgical preparation is an important decrease in hepatic and testicular volume. Different sham-operation methods including a laparotomy were used as controls in each case. Given that the liver volume varies greatly in comparison to body weight in the sham-operated animals, this paper aims to evaluate the possible consequences of the sham operation. It concludes that control animals without manipulation, in addition to the respective controls of portacaval shunt, should be used in every case.


Descriptive sensory analysis of Aceto Balsamico Tradizionale di Modena DOP and Aceto Balsamico Tradizionale di Reggio Emilia DOP.
Aceto Balsamico Tradizionale (ABT) is a typical Italian vinegar available in two different forms: Aceto Balsamico Tradizionale di Modena DOP (ABTM) and Aceto Balsamico Tradizionale di Reggio Emilia DOP (ABTRE). ABT is obtained by alcoholic fermentation and acetic bio-oxidation of cooked grape must and aged at least 12 years in wooden casks and is known and sold around the world. Despite this widespread recognition, data on sensory characteristics of these products are very scarce. Therefore a descriptive analysis was conducted to define a lexicon for the ABT sensory profile and to create a simple, stable and reproducible synthetic ABT for training panellists. A lexicon of 20 sensory parameters was defined and validated and a synthetic ABT was prepared as standard reference. Simple standards for panellist training were also defined and the sensory profiles of ABTM and ABTRE were obtained. The obtained results confirm that descriptive analysis can be used for the sensory characterisation of ABT and that the sensory profiles of ABTM and ABTRE are very different. Furthermore, the results demonstrate that a lexicon and proper standard references are essential for describing the sensory qualities of ABT both for technical purposes and to protect the product from commercial fraud.


Modulation of immunogenicity and immunoprotection of mucosal vaccine against coxsackievirus B3 by optimizing the coadministration mode of lymphotactin adjuvant.
Induction of potent mucosal immune response is a goal of current vaccine strategies against mucus-infectious pathogens such as Coxsackievirus B3 type (CVB3). We previously showed that administration of lymphotactin (LTN) as an adjuvant could enhance the specific immune responses against a mucosal gene vaccine, chitosan-pVP1, against CVB3. To optimize the coadministration mode of the mucosal adjuvant, we compared the mucosal immune responses induced by chitosan-DNA vaccine with different combinations of the target VP1 antigen gene and the adjuvant LTN gene. The two genes were either cloned in separate vectors or coexpressed as a fusion or bicistron protein in the same vector before encapsulation in chitosan nanoparticles. Four doses of various adjuvant-combined chitosan-DNA were intranasally administrated to mice before challenge with CVB3. The results indicated that chitosan-formulated pVP1-LTN fusion plasmid exhibited very weak improvement of CVB3-specific immune responses. Although the bicistronic coexpression of LTN with VP1 was expected to be powerful, this combination had enhanced effects on serum IgG and systemic T cell immune responses, but not on mucosal T cell immunity. Coimmunization with VP1 and LTN as separate chitosan-DNA formulation remarkably enhanced antibody and T cell immune responses both in systemic and mucosal immune compartments, leading to the most desirable preventive effect on viral myocarditis. Taken together, how the adjuvant is combined with the target antigen has a strong influence on the mucosal immune responses induced by mucosal DNA vaccines.


The level of performance stabilization influences motor adaptation on an isometric force control task.
This study compared the effects of two levels of performance stabilization on the adaptation to unpredictable perturbations in an isometric control force task with the goal of controlling 40% of the maximum force. The experiment consisted of pre-exposure and exposure phases. In the pre-exposure two levels of performance stabilization were manipulated: a stabilization group (SG) performed three trials in a row while maintaining 40% of the maximum force for three seconds and an absolute error less than or equal to 5% (i.e., the criteria of performance), and a superstabilization group (SSG) performed six blocks of the same criteria. During the exposure phase, the task was the same as that in the pre-exposure phase; however, it was inserted 9 trials of perturbations when the task goal changed to 60% of the maximum force. We measured the %RMSE, RMS from the biceps and triceps brachii and co-contraction. In the pre-exposure phase, both groups showed similar performance and muscle activity. When exposed to the perturbations, SSG performance more quickly returned to the previous level of accuracy, showed lower muscle activation and demonstrated a greater muscle co-contraction than did SG. The results give support to the adaptive process model on motor learning.


Lacticaseibacillus rhamnosus GG inhibits infection of human keratinocytes by Staphylococcus aureus through mechanisms involving cell surface molecules and pH reduction.
Beneficial bacteria represent an emerging tool against topical diseases, including infection caused by Staphylococcus aureus. Here, we investigated several anti-pathogenic mechanisms of the model probiotic Lacticaseibacillus rhamnosus GG against a clinical S. aureus isolate by implementing various mutants lacking important cell surface molecules. We analysed adhesion of L. rhamnosus and competitive adhesion with S. aureus to primary human keratinocytes, L. rhamnosus and S. aureus auto- and co-aggregation, S. aureus growth inhibition, keratinocyte viability increase, and monocyte Toll-like receptor (TLR) activation by L. rhamnosus as such, or with S. aureus. L. rhamnosus mutated in SpaCBA pili exhibited reduced adhesion to keratinocytes, reduced ability to prevent S. aureus adhesion to keratinocytes and reduced co-aggregation with S. aureus. Mutants in cell wall exopolysaccharides showed enhanced adhesion to keratinocytes and TLR activation in monocytes, suggesting involvement of additional cell surface molecules masked by exopolysaccharides. All L. rhamnosus strains inhibited S. aureus growth, likely due to acidification of the medium. Live (but not UV-inactivated) L. rhamnosus significantly reduced inflammatory TLR activation in monocytes by S. aureus. These data suggest the key role of SpaCBA pili and additional contribution of other cell surface molecules as well as secreted components of L. rhamnosus GG in the multifactorial inhibition of S. aureus adhesion and toxicity in the skin niche.


Addressing the Complexity of Tourette's Syndrome through the Use of Animal Models.
Tourette's syndrome (TS) is a neurodevelopmental disorder characterized by fluctuating motor and vocal tics, usually preceded by sensory premonitions, called premonitory urges. Besides tics, the vast majority-up to 90%-of TS patients suffer from psychiatric comorbidities, mainly attention deficit/hyperactivity disorder (ADHD) and obsessive-compulsive disorder (OCD). The etiology of TS remains elusive. Genetics is believed to play an important role, but it is clear that other factors contribute to TS, possibly altering brain functioning and architecture during a sensitive phase of neural development. Clinical brain imaging and genetic studies have contributed to elucidate TS pathophysiology and disease mechanisms; however, TS disease etiology still is poorly understood. Findings from genetic studies led to the development of genetic animal models, but they poorly reflect the pathophysiology of TS. Addressing the role of neurotransmission, brain regions, and brain circuits in TS disease pathomechanisms is another focus area for preclinical TS model development. We are now in an interesting moment in time when numerous innovative animal models are continuously brought to the attention of the public. Due to the diverse and largely unknown etiology of TS, there is no single preclinical model featuring all different aspects of TS symptomatology. TS has been dissected into its key symptomst hat have been investigated separately, in line with the Research Domain Criteria concept. The different rationales used to develop the respective animal models are critically reviewed, to discuss the potential of the contribution of animal models to elucidate TS disease mechanisms.


Characterization of CYP76M5-8 indicates metabolic plasticity within a plant biosynthetic gene cluster.
Recent reports have revealed genomic clustering of enzymatic genes for particular biosynthetic pathways in plant specialized/secondary metabolism. Rice (Oryza sativa) carries two such clusters for production of antimicrobial diterpenoid phytoalexins, with the cluster on chromosome 2 containing four closely related/homologous members of the cytochrome P450 CYP76M subfamily (CYP76M5-8). Notably, the underlying evolutionary expansion of these CYP appears to have occurred after assembly of the ancestral biosynthetic gene cluster, suggesting separate roles. It has been demonstrated that CYP76M7 catalyzes C11α-hydroxylation of ent-cassadiene, and presumably mediates an early step in biosynthesis of the derived phytocassane class of phytoalexins. Here we report biochemical characterization of CYP76M5, -6, and -8. Our results indicate that CYP76M8 is a multifunctional/promiscuous hydroxylase, with CYP76M5 and -7 seeming to provide only redundant activity, while CYP76M6 seems to provide both redundant and novel activity, relative to CYP76M8. RNAi-mediated double knockdown of CYP76M7 and -8 suppresses elicitor inducible phytocassane production, indicating a role for these monooxygenases in phytocassane biosynthesis. In addition, our data suggests that CYP76M5, -6, and -8 may play redundant roles in production of the oryzalexin class of phytoalexins as well. Intriguingly, the preceding diterpene synthase for oryzalexin biosynthesis, unlike that for the phytocassanes, is not found in the chromosome 2 diterpenoid biosynthetic gene cluster. Accordingly, our results not only uncover a complex evolutionary history, but also further suggest some intriguing differences between plant biosynthetic gene clusters and the seemingly similar microbial operons. The implications for the underlying metabolic evolution of plants are then discussed.


Michigan Marijuana Legalization: Correlations Among Cannabis Use, Mental Health, and Other Factors.
There are health implications with the statewide legalization of recreational marijuana that are still not fully understood and require further examination. This study evaluates the prevalence of marijuana use in patients being treated for a variety of conditions and whether correlations exist between marijuana use, mental health conditions, and concomitant use of psychotropic medications. Data were collected from an electronic medical record (EMR) as part of a retrospective chart audit. A total of 500 charts were reviewed during a six-month timeframe from December 1, 2018 to May 31, 2019 with the start date approximating the timing of when marijuana became recreationally legalized in the State of Michigan. This study demonstrated a point prevalence of 15.8% since 79 of the 500 charts reviewed had marijuana use documented. Additionally, marijuana users were more likely to have a history of cocaine use, schizophrenia, antipsychotic use, and tobacco use. Trends identified in this study provide a comparison point for the local prevalence of marijuana use immediately post state-wide legalization, with a projected increasing trend due to the removal of legal barriers.


Subarachnoid hemorrhage from multiple neurofibromas of the cauda equina: case report.
The authors report a case of spinal subarachnoid hemorrhage caused by neurofibromas of the cauda equina. An examination of the literature revealing 13 similar cases and an analysis of this case showing three neurofibromas of the cauda equina prompted an hypothesis emphasizing the origin of bleeding and the difficulties associated with a correct diagnosis.


Familial benign pemphigus atypical localization.
We present an atypical case of familial benign pemphigus (Hailey-Hailey disease), which presented as crusted, annular plaques limited to the back without intertriginous involvement. We could not find in the literature another patient with plaques located solely on the back without a prior history of classical disease.


With a Little Help from My Friends: Microbial Partners in Integrative and Comparative Biology-An Introduction to the Symposium.
The role that host-associated microbes play in animal biology is gaining attention in comparative biology. Numerous research groups study the roles that microbes play in human health and nutrition, or in enhancing the production of agricultural animals. However, inclusion of host-associated microbes into research questions of integrative and comparative biology has lagged behind. We hosted a symposium to bring together top researchers in the field of host-associated microbes who also incorporate aspects of integrative and comparative biology. In this introduction, we highlight recent research demonstrating the profound roles that host-associated microbes play in many aspects of animal biology, such as immune function, endocrinology, and even behavior. It is our hope that integrative and comparative biologists will begin to include aspects of host-associated microbes into their research programs, enhancing both the fields of comparative biology and host-microbe interactions.


The Ionophores CCCP and Gramicidin but Not Nigericin Inhibit Trypanosoma brucei Aquaglyceroporins at Neutral pH.
Human African trypanosomiasis (HAT) is caused by Trypanosoma brucei parasites. The T. brucei aquaglyceroporin isoform 2, TbAQP2, has been linked to the uptake of pentamidine. Negative membrane potentials and transmembrane pH gradients were suggested to promote transport of the dicationic antitrypanosomal drug. Application of ionophores to trypanosomes further hinted at direct inhibition of TbAQP2 by carbonyl cyanide m-chlorophenyl hydrazone (CCCP). Here, we tested for direct effects of three classical ionophores (CCCP, nigericin, gramicidin) on the functionality of TbAQP2 and the related TbAQP3 at conditions that are independent from the membrane potential or a proton gradient. We expressed TbAQP2 and TbAQP3 in yeast, and determined permeability of uncharged glycerol at neutral pH using stopped-flow light scattering. The mobile proton carrier CCCP directly inhibited TbAQP2 glycerol permeability at an IC50 of 2 µM, and TbAQP3 to a much lesser extent (IC50 around 1 mM) likely due to different selectivity filter layouts. Nigericin, another mobile carrier, left both isoforms unaffected. The membrane-integral pore-forming gramicidin evenly inhibited TbAQP2 and TbAQP2 in the double-digit micromolar range. Our data exemplify the need for suitable controls to detect unwanted ionophore side effects even when used at concentrations that are typically recommended to disturb the transmembrane ion distribution.


Malignant teratoid medulloepithelioma with brain and kidney involvement in a dog.
A tumor of the left eye with involvement of the brain and kidney was diagnosed in a 4-year-old Neapolitan mastiff. The dog presented with acute glaucoma of the left eye. Peripheral corneal vascularization and severe corneal edema obscured examination of deeper structures. Because of concurrent progressive neurologic signs the dog was euthanized and a postmortem examination was performed. The eyes, brain and samples from the lung, heart, liver and kidneys were fixed in 4% neutral buffered formalin and embedded in paraffin wax. Sections were examined by light microscopy including histochemical and immunohistochemical staining. The ocular tumor originated from the ciliary body and was composed of small islets and cords of poorly differentiated, oval to polyhedral cells surrounded by abundant ground substance. Areas with chondroid differentiation were observed. Rosettes were not found. Metastases with the same morphology were present in the brain and in one kidney. An intraocular malignant teratoid medulloepithelioma with metastases to the brain and kidney is presented.


Phase II Study of Nonmyeloablative Allogeneic Bone Marrow Transplantation for B Cell Lymphoma with Post-Transplantation Rituximab and Donor Selection Based First on Non-HLA Factors.
Outcomes of nonmyeloablative (NMA), HLA-haploidentical (haplo), related-donor allogeneic blood or marrow transplantation (allo-BMT) with high-dose post-transplantation cyclophosphamide (PTCy) appear to be similar to those using HLA-matched donors. Thus, it may be possible to prioritize donor factors other than HLA matching that could enhance antitumor activity. The Fc receptor polymorphism FCGR3A-158VV may confer greater sensitivity to rituximab than FCGR3A-158FF. In a prospective phase II study of NMA, related-donor allo-BMT with PTCy and post-transplantation rituximab for patients with B cell lymphomas, we hypothesized that donor selection that prioritized FCGR3A-158 polymorphism over HLA matching would be feasible, safe, and improve outcomes. The primary endpoint was 1-year progression-free survival (PFS). Of 83 patients transplanted (median age, 59 years), 69 (83%) received haplo grafts. Fifty-four (65%) received a graft that maintained or improved their Fc receptor polymorphism status. With 2.6-year median follow-up, the 1-year PFS and overall survival (OS) probabilities were 71% and 86%, respectively, with 1-year relapse and nonrelapse mortality (NRM) probabilities of 20% and 8%. At 1 year, the probability of acute grades II to IV graft-versus-host disease (GVHD) was 41%, with acute grades III to IV GVHD probability of 5% and chronic GVHD probability of 11%. Among haplo transplants, the 1-year probabilities of PFS, OS, relapse, and NRM were 70%, 83%, 20%, and 10%, respectively. No differences in outcomes were observed based on donor FCGR3A-158 polymorphism. Excess infection risk was not apparent with post-transplantation rituximab. Although donor selection based on FCGR3A-158 polymorphism was not shown to influence PFS, this study suggests that donor selection based on criteria other than best HLA match is feasible and safe. This study opens the way for the future investigation of donor prioritization based on promising non-HLA factors that may improve antitumor activity and decrease relapse after allo-BMT. This study was registered at www.clinicaltrials.gov as NCT00946023.


Understanding the relations between RAN letter subtest components and word reading in first-grade students.
First-grade students (N = 221) were individually tested on a battery of cognitive and achievement measures of verbal fluency, visual attention, phonological awareness, orthographic recognition, rapid automatized naming (RAN) of letters and objects, and reading. All tests were subjected to postacquisition scoring, and all RAN measures were segregated into measures of articulation time, pause time, and consistency of the pause time. Structural equation modeling demonstrated that word reading was directly and significantly predicted by RAN letter naming and general RAN cognitive processing time of objects. Moreover, RAN letter reading constructs were significantly and directly predicted by the latent variables of phonological awareness, orthographic recognition, and general RAN object articulation and cognitive processing times. RAN letter naming constructs were also significantly and indirectly predicted by visual attention. The reading model was found to be consistent with a total mediation of the relation of phonological awareness and reading through RAN letter naming and supported the validity of the RAN letter naming subtest as a basic letter reading test. These findings supported the double-deficit hypothesis for letter reading. We suggest that phonological memory is a basic factor underlying general RAN cognitive processing time of objects and domain-specific information associated with phonemes and their graphic representations.


Ruminal Degradation of Rumen-Protected Glucose Influences the Ruminal Microbiota and Metabolites in Early-Lactation Dairy Cows.
Rumen-protected glucose (RPG) plays an important role in alleviating the negative energy balance of dairy cows. This study used a combination of rumen microbes 16S and metabolomics to elucidate the changes of rumen microbial composition and rumen metabolites of different doses of RPG's rumen degradation part in early-lactation dairy cows. Twenty-four multiparous Holstein cows in early lactation were randomly allocated to control (CON), low-RPG (LRPG), medium-RPG (MRPG), or high-RPG (HRPG) groups in a randomized block design. The cows were fed a basal total mixed ration diet with 0, 200, 350, and 500 g of RPG per cow per day, respectively. Rumen fluid samples were analyzed using Illumina MiSeq sequencing and ultrahigh-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry. MRPG supplementation increased bacterial richness and diversity, including increasing the relative abundance of cellulolytic bacteria, such as Ruminococcus, Lachnospiraceae_NK3A20_group, Ruminiclostridium, and Lachnospiraceae_UCG-008 MRPG significantly increased the concentrations of acetate, propionate, butyrate, and total volatile fatty acid in the rumen. Ruminal fluid metabolomics analysis showed that RPG supplementation could significantly regulate the synthesis of amino acids digested by protozoa in the rumen. Correlation analysis of the ruminal microbiome and metabolome revealed some potential relationships between major bacterial abundance and metabolite concentrations. Our analysis found that RPG supplementation of different doses can change the diversity of microorganisms in the rumen and affect the rumen fermentation pattern and microbial metabolism and that a daily supplement of 350 g of RPG might be the ideal dose.IMPORTANCE Dairy cows in early lactation are prone to a negative energy balance because their dry matter intake cannot meet the energy requirements of lactation. Rumen-protected glucose is used as an effective feed additive to alleviate the negative energy balance of dairy cows in early lactation. However, one thing that is overlooked is that people often think that rumen-protected glucose is not degraded in the rumen, thus ignoring its impact on the microorganisms in the rumen environment. Our investigation and previous experiments have found that rumen-protected glucose is partially degraded in the rumen. However, there are few reports on this subject. Therefore, we conducted research on this problem and found that rumen-protected glucose supplementation at 350 g/day can promote the development and metabolism of rumen flora. This provides a theoretical basis for the extensive application of rumen bypass glucose at a later stage.


Deletion mutagenesis of p22phox subunit of flavocytochrome b558: identification of regions critical for gp91phox maturation and NADPH oxidase activity.
The heterodimeric flavocytochrome b558, comprised of the two integral membrane proteins p22phox and gp91phox, mediates the transfer of electrons from NADPH to molecular oxygen in the phagocyte NADPH oxidase to generate the superoxide precursor of microbicidal oxidants. This study uses deletion mutagenesis to identify regions of p22phox required for maturation of gp91phox and for NADPH oxidase activity. N-terminal, C-terminal, or internal deletions of human p22phox were generated and expressed in Chinese hamster ovary cells with transgenes for gp91phox and two other NADPH oxidase subunits, p47phox, and p67phox. The results demonstrate that p22phox-dependent maturation of gp91phox carbohydrate, cell surface expression of gp91phox, and the enzymatic function of flavocytochrome b558 are closely correlated. Whereas the 5 N-terminal and 25 C-terminal amino acids are dispensable for these functions, the N-terminal 11 amino acids of p22phox are required, as is a hydrophilic region between amino acids 65 and 90. Upon deletion of 54 residues at the C terminus of p22phox (amino acids 142-195), maturation and cell surface expression of gp91phox was still preserved, although NADPH oxidase activity was absent, as expected, due to removal of a proline-rich domain between amino acids 151-160 that is required for recruitment of p47phox. Antibody binding studies indicate that the extreme N terminus of p22phox is inaccessible in the absence of cell permeabilization, supporting a model in which both the N- and C-terminal domains of p22phox extend into the cytoplasm, anchored by two membrane-embedded regions.


Comparative Label-Free Quantitative Proteomics Analysis Reveals the Essential Roles of N-Glycans in Salt Tolerance by Modulating Protein Abundance in Arabidopsis.
Many pieces of evidence show that the adaptive response of plants to salt stress requires the maturation of N-glycan on associated proteins. However, it is still little known about the salt-responsive glycoproteins that function in this process. In the present study, we identified salt-responsive glycoproteins in wild-type (WT) Arabidopsis and two mutants defective in N-glycan maturation, mns1 mns2 and cgl1. A total of 97 proteins with abundance changes of >1.5- or <0.67-fold were identified against salt stress by label-free liquid chromatography coupled mass spectrometry (LC-MS/MS) quantitative analyses. A comparison of differentially abundant glycoproteins (DAGs) indicated the substrate preferences regulated by MNS1/MNS2 and CGL1. In addition, the DAGs in mns1 mns2 hardly form functional regulatory networks in STRING analysis. Comparably, the regulatory network in cgl1 was visible and shared overlapping with that in WT. Such difference may supply the evidence to partially explain the lower salt sensitivity of mutant cgl1 than mns1 mns2. We further confirmed that two N-glycosylation clients, peroxidases PRX32 and PRX34, were involved in the salt stress response since the double mutants showed enhanced salt sensitivity. Together, our study provided proteomic evidence that N-glycans are crucial for modulating stress-responsive protein levels, and several novel glycoproteins responsible for salt stress tolerance in Arabidopsis were listed. Data are available via ProteomeXchange with identifier PXD006893.


C60-β-cyclodextrin conjugates for enhanced nucleus delivery of doxorubicin.
We demonstrate the use of water-soluble C60-β-cyclodextrin conjugates to encapsulate and deliver doxorubicin to the cell nucleus. The behaviour of the fullerene aggregates inside cells is dictated by the functionalization of the C60 cage. While both the C60 conjugates are taken up by lysosomes upon cellular entry, only the one with a hydroxylated cage rapidly escaped the lysosome. The drug delivery system (DDS) with a hydroxylated C60 cage showed significantly enhanced doxorubicin delivery to the cell nucleus, whereas the DDS with a hydrophobic C60 cage was trapped in the lysosome for a longer time and showed significantly reduced doxorubicin delivery to the nucleus. This study opens new paths towards advanced fullerene-based DDSs for small molecule drugs.


Bone tissue modelling and remodelling following guided bone regeneration in combination with biphasic calcium phosphate materials presenting different microporosity.
The aim of this study was to investigate bone regeneration following application of a novel biphasic calcium phosphate (BCP I) composed of microstructured granules of 90% β-tricalcium phosphate (β-TCP)/10% hydroxyapatite (HA) compared to BCP non-microstructured biphasic calcium phosphate with a composite of 60% hydroxyapatite/40% β-TCP (BCP II) and a deproteinized bovine bone mineral (DBBM) at surgically created defects in the mandible of minipigs in a combined approach with guided bone regeneration (GBR). Sixteen minipigs were used for the study. Lower premolars P2, P3, P4 and first molar M1 were extracted. Following 3 months of healing, two defects with a width and depth of 7 mm were created bilaterally in the mandible. The different grafting materials were randomly placed in the created defects and covered by means of a collagen membrane. After 3 and 8 weeks, biopsies were sampled. All specimens were evaluated with descriptive histology and histomorphometric evaluations complemented by micro-CT scan analysis. All three biomaterials presented with higher bone volume at 8 weeks compared to 3 weeks (P < 0.0442). BCP I and DBBM demonstrated a significant higher amount of bone formation compared to BCP II at 8 weeks (P < 0.0328). BCP I also demonstrated a significant higher percentage of remaining graft volume compared to the other test groups both at 3 and 8 weeks (P < 0.0001 to P < 0.0003). Congruently, defects containing BCP I showed a significant higher amount of mineralized tissue compared to the other groups. All the three test materials performed well with regard to bone formation at 8 weeks. BCP I showed significant higher amounts of newly formed bone despite a higher remaining graft volume compared to the other groups. With regard to the regenerative outcome, all the three materials can be recommended for clinical use.


Predictors of long-term abstinence among Chinese smokers following treatment: the role of personality traits.
Four hundred and thirty-five Chinese smokers who had attended the first smoking cessation clinic and received stage-matched individualized cessation counseling in Hong Kong during the period of August 2000 to January 2002 were successfully followed-up between February and August 2008. Some 38% of the participants (165/495) had stopped smoking during the 12-month follow-up after treatment. Participants with higher conscientiousness score (OR = 1.65, 95% CI: 1.09 - 2.48) and lower openness to experience score (OR = 0.64, 95% CI: 0.45 - 0.92) showed a greater likelihood of quitting smoking after controlling the effect of daily cigarettes smoked and stage of readiness at baseline. The study suggests an important role of conscientiousness and openness to experience on long-term quitting behaviors following treatment, and provides useful information for the development of matched intervention for smoking cessation among Chinese smokers.


Towards stationary phases for chromatography on a microchip: molded porous polymer monoliths prepared in capillaries by photoinitiated in situ polymerization as separation media for electrochromatography.
Photoinitiated free radical polymerization has been used for the preparation of porous polymer monoliths within UV transparent fused silica capillaries and quartz tubes. These formats were used as models for the preparation of the separation media within channels of microfabricated devices. A mixture of ethylene dimethacrylate, butyl methacrylate, and 2-acrylamido-2-methyl-1-propanesulfonic acid was polymerized in the presence of a porogenic solvent consisting of 1-propanol, 1,4-butanediol, and water at room temperature under UV irradiation. Modification of the porogen composition enables the tailoring of pore size within the broad range from ca. 100 to 4000 nm. Scanning electron micrographs confirmed the homogeneity of the porous structure of the materials prepared, even in a quartz tube with a diameter as large as 4 mm. Separation properties of the resulting capillary columns were tested in capillary electrochromatography (CEC) mode using a mixture of thiourea and eight aromatic compounds. Plate number as high as 210 000 plates/m were found for a capillary column with optimized porous properties. The monolithic columns were also able to separate mixtures of peptides.


Is understanding regret dependent on developments in counterfactual thinking?
Children's understanding of counterfactual emotions such as regret and relief develops relatively late compared to their ability to imagine counterfactual worlds. We tested whether a late development in counterfactual thinking: understanding counterfactuals as possibilities, underpinned children's understanding of regret. Thirty 5- and 6-year-olds completed tasks assessing counterfactual thinking and understanding regret. Performance on the counterfactual task was better than that on the regret task. We suggest that thinking about counterfactuals as possibilities is a necessary but not sufficient cognitive development in children's understanding of regret. We discuss how other developments in counterfactual thinking may underpin children's emotional understanding.


Clinicians' perspectives on the implementation of patient decision aids in the emergency department: A qualitative interview study.
Decision aids (DAs) are tools to facilitate and standardize shared decision making (SDM). Although most emergency clinicians (ECs) perceive SDM appropriate for emergency care, there is limited uptake of DAs in clinical practice. The objective of this study was to explore barriers and facilitators identified by ECs regarding the implementation of DAs in the emergency department (ED). We conducted a qualitative interview study guided by implementation science frameworks. ECs participated in interviews focused on the implementation of DAs for the disposition of patients with low-risk chest pain and unexplained syncope in the ED. Interviews were recorded and transcribed verbatim. We then iteratively developed a codebook with directed qualitative content analysis. We approached 25 ECs working in urban New York, of whom 20 agreed to be interviewed (mean age, 41 years; 25% women). The following 6 main barriers were identified: (1) poor DA accessibility, (2) concern for increased medicolegal risk, (3) lack of perceived need for a DA, (4) patient factors including lack of capacity and limited health literacy, (5) skepticism about validity of DAs, and (6) lack of time to use DAs. The 6 main facilitators identified were (1) positive attitudes toward SDM, (2) patient access to follow-up care, (3) potential for improved patient satisfaction, (4) potential for improved risk communication, (5) strategic integration of DAs into the clinical workflow, and (6) institutional support of DAs. ECs identified multiple barriers and facilitators to the implementation of DAs into clinical practice. These findings could guide implementation efforts targeting the uptake of DA use in the ED.


The potential role of c-MYC and polyamine metabolism in multiple drug resistance in bladder cancer investigated by metabonomics.
Bladder cancer has a high incidence worldwide accompanies by high recurrent rate after treatment. The emergence of primary or acquired chemotherapy resistance leads to poor efficacy in many cases. To explore the underlying mechanisms of drug resistance, we firstly established a drug-resistant cell model T24/THP by repeated exposure of T24 cells to pirarubicin (THP) whose concentration increases gradually. Non-targeted metabolomics was performed to identify metabolic changes and key metabolism pathways variance in T24/THP cells. Pathway enrichment analysis demonstrated that the arginine and proline metabolic pathway was the most significantly changed pathway, where two representative members of polyamine, putrescine and spermidine were remarkably down regulated in T24/THP. Subsequent experiments further confirmed that ornithine decarboxylase (ODC1) and spermidine synthase (SRM), the key enzymes involved in the synthesis of these compounds, also showed a stable low expression in T24/THP. However, knocking down of ODC1 and SRM sensitized cells to chemotherapy treatment while overexpression of these two enzymes enhances chemotherapy resistance. This leaded to the point that ODC1 and SRM themselves are more likely to promote the drug resistance, which appears to contradict their low expression in T24/THP. We hypothesize that their diminished levels were due to the declined activity of genes upstream. According to this line of thought, we found that c-MYC was also down-regulated in T24/THP and its content could be significantly affected by drug administration. In addition, c-MYC could not only regulate the expression levels of ODC1 and SRM but also influence drug resistance in T24/THP. In conclusion, alterations in gene expression of ODC1 and SRM in drug resistance cell line is probably mediated by some upstream regulators rather than antineoplastic agents alone. Exploration of upstream signals and research on detailed regulatory mechanism, thereby understanding the actual role of c-MYC and polyamine in response to chemotherapy, can become a potential field direction to overcome drug resistance in bladder cancer.


Increased Shamal winds and dust activity over the Arabian Peninsula during the COVID-19 lockdown period in 2020.
While anthropogenic pollutants have decreased during the lockdown imposed as an effort to contain the spread of the Coronavirus disease 2019 (COVID-19), changes in particulate matter (PM) do not necessarily exhibit the same tendency. This is the case for the eastern Arabian Peninsula, where in March-June 2020, and with respect to the same period in 2016-2019, a 30 % increase in PM concentration is observed. A stronger than normal nocturnal low-level jet and subtropical jet over parts of Saudi Arabia, in response to anomalous convection over the tropical Indian Ocean, promoted enhanced and more frequent episodes of Shamal winds over the Arabian Peninsula. Increased surface winds associated with the downward mixing of momentum to the surface fostered, in turn, dust lifting and increased PM concentrations. The stronger low-level winds also favoured long-range transport of aerosols, changing the PM values downstream. The competing effects of reduced anthropogenic and increased dust concentrations leave a small positive signal (<5 W m-2) in the net surface radiation flux (Rnet), with the former dominating during daytime and the latter at night. However, in parts of the Arabian Gulf, Sea of Oman and Iran Rnet increased by >20 W m-2 with respect to the baseline period, owing to a clearer environment and weaker winds. It is concluded that a reduction in anthropogenic emissions due to the lockdown does not necessarily go hand in hand with lower particulate matter concentrations. Therefore, emissions reduction strategies need to account for feedback effects in order to reach the planned long-term outcomes.


Diffusion and perfusion magnetic resonance imaging in brain tumors.
The article reviews recent progress made in the field of diffusion and perfusion magnetic resonance (MR) imaging and presents possible applications in brain tumors. Diffusion, a new parameter, provides useful data to assess tissue structure and function. Perfusion MR imaging gives results that are somewhat similar to those obtained with classic non-MR imaging methods, but it offers several potential advantages.


Cooccurrence of Five Pathogenic Legionella spp. and Two Free-Living Amoebae Species in a Complete Drinking Water System and Cooling Towers.
Pathogenic Legionella species grow optimally inside free-living amoebae to concentrations that increase risks to those who are exposed. The aim of this study was to screen a complete drinking water system and cooling towers for the occurrence of Acanthamoeba spp. and Naegleria fowleri and their cooccurrence with Legionella pneumophila, Legionella anisa, Legionella micdadei, Legionella bozemanii, and Legionella longbeachae. A total of 42 large-volume water samples, including 12 from the reservoir (water source), 24 from two buildings (influents to the buildings and exposure sites (taps)), and six cooling towers were collected and analyzed using droplet digital PCR (ddPCR). N. fowleri cooccurred with L. micdadei in 76 (32/42) of the water samples. In the building water system, the concentrations of N. fowleri and L. micdadei ranged from 1.5 to 1.6 Log10 gene copies (GC)/100 mL, but the concentrations of species increased in the cooling towers. The data obtained in this study illustrate the ecology of pathogenic Legionella species in taps and cooling towers. Investigating Legionella's ecology in drinking and industrial waters will hopefully lead to better control of these pathogenic species in drinking water supply systems and cooling towers.


Vertically oriented MoS2 nanoflakes coated on 3D carbon nanotubes for next generation Li-ion batteries.
The advent of advanced electrode materials has led to performance enhancement of traditional lithium ion batteries (LIBs). We present novel binder-free MoS2 coated three-dimensional carbon nanotubes (3D CNTs) as an anode in LIBs. Scanning transmission electron microscopy analysis shows that vertically oriented MoS2 nanoflakes are strongly bonded to CNTs, which provide a high surface area and active electrochemical sites, and enhanced ion conductivity at the interface. The electrochemical performance shows a very high areal capacity of ~1.65 mAh cm-2 with an areal density of ~0.35 mg cm-2 at 0.5 C rate and coulombic efficiency of ~99% up to 50 cycles. The unique architecture of 3D CNTs-MoS2 is indicative to be a promising anode for next generation Li-ion batteries with high capacity and long cycle life.


Impact of allogenic and autologous transfusion on immune function in patients with tumors.
To observe the effects of allogeneic and autologous transfusion on cellular immunity, humoral immunity and secretion of serum inflammatory factors and perforin during the perioperative period in patients with malignant tumors. A total of 80 patients (age: 38-69 years; body weight: 40-78 kg; ASA I - II) receiving radical operation for gastro-intestinal cancer under general anesthesia were selected. All the patients were divided into four groups based on the methods of infusion and blood transfusion: blank control group (Group C), allogeneic transfusion group (group A), hemodiluted autotransfusion Group (Group H) and hemodiluted autotransfusion + allogenic transfusion Group (A+H group). Venous blood was collected when entering into the surgery room (T0), immediately after surgery (T1) and 24h (T2), 3d (T3) and 7d (T4) after surgery, respectively. Moreover, flow cytometry was applied to assess changes of peripheral blood T cell subpopulations and NK cells. Enzyme linked immunosorbent assays were performed to determine levels of IL-2, IL-10, TNF-α and perforin. Immune turbidimetry was employed to determine the changes in serum immunoglobulin. Both CD3+ and NK cells showed a decrease at T1 and T2 in each group, among which, in group A, CD3+ decreased significantly at T2 (P<0.05) compared with other groups, and CD3+ and NK cell reduced obviously only in group A at T3 and T4 (P<0.05). CD4+ cells and the ratio of D4+/CD8+ were decreased in groups A, C and A+H at T1 and T2 (P<0.05). No significant intra- and inter-group differences were observed in CD8+ of the four groups (P<0.05). IL-2 declined in group C at T1 and T2 (P<0.05) and showed a decrease in group A at each time point (P<0.05). Moreover, IL-2 decreased in group A + H only at T1. No significant difference was found in each group at T1 (P<0.05). More significant decrease in group C at T2, T3 and T4 compared with group A (P<0.05), and there were no significant differences among other groups (P>0.05). IL-10 increased at T1 and T2 in each group (P<0.05), in which it had an obvious increase in group A, and increase of IL-10 occurred only in group A at T3 and T4 (P<0.05). TNF-α level rose at T1 (P<0.05), no inter- and intra-group difference was found in perforin in all groups (P<0.05). Compared with the preoperation, both IgG and IgA level decreased at T1 in each group (P<0.05), and they declined only in Group A at T2 and T3 (P<0.05), and these parameters were back to the preoperative levels in other groups. No significant differences were observed between preoperative and postoperative IgG and IgA levels in each group at T4 (P>0.05). No obvious inter- and intra-group changes were found in IgM in the four groups (P>0.05). Allogeneic transfusion during the perioperative period could obviously decrease the number of T cell subpopulations and NK cells and the secretion of stimulating cytokines and increase the secretion of inhibiting cytokines in patients with malignant tumors, thus causing a Th1/Th2 imbalance and transient decreasing in the content of plasma immune globulin. Autologous transfusion has little impact and may even bring about some improvement of postoperative immune function in patients with tumors. Therefore, cancer patients should receive active autologous transfusion during the perioperative period in place of allogeneic transfusion.


Hematologic parameters during treatment with high-dose medroxyprogesterone acetate.
Blood clotting, platelet aggregation, complete blood count and lipid profile were evaluated in 12 postmenopausal patients with advanced breast cancer. Patients under treatment with high-dose MPA were considered not at risk for thomboembolic disease and were given MPA orally, 800 mg/day, for at least 3 months. Laboratory investigations were performed prior to treatment with MPA then once weekly during the first month and every 2 weeks during the following months. PTT, TEG, antithrombin III and platelet adhesiveness underwent statistically significant changes, tending towards hypercoagulability, although, on the average, they did not exceed the upper normal range. The authors conclude that a clinically relevant thrombotic activity cannot be attributed to MPA at the administered oral doses in the absence of additional risk factors.


In Situ Transmission Electron Microscopy to Study the Location and Distribution Effect of Pt on the Reduction of Co3 O4 -SiO2.
The addition of Pt generally promotes the reduction of Co3 O4 in supported catalysts, which further improves their activity and selectivity. However, due to the limited spatial resolution, how Pt and its location and distribution affect the reduction of Co3 O4 remains unclear. Using ex situ and in situ ambient pressure scanning transmission electron microscopy, combined with temperature-programmed reduction, the reduction of silica-supported Co3 O4 without Pt and with different location and distribution of Pt is studied. Shrinkage of Co3 O4 nanoparticles is directly observed during their reduction, and Pt greatly lowers the reduction temperature. For the first time, the initial reduction of Co3 O4 with and without Pt is studied at the nanoscale. The initial reduction of Co3 O4 changes from surface to interface between Co3 O4 and SiO2 . Small Pt nanoparticles located at the interface between Co3 O4 and SiO2 promote the reduction of Co3 O4 by the detachment of Co3 O4 /CoO from SiO2 . After reduction, the Pt and part of the Co form an alloy with Pt well dispersed. This study for the first time unravels the effects of Pt location and distribution on the reduction of Co3 O4 nanoparticles, and helps to design cobalt-based catalysts with efficient use of Pt as a reduction promoter.


Evaluation of Male Infertility Prevalence with Clinical Outcomes in Middle Anatolian Region.
the aim of this study is to determine the prevalence of male factor infertility with the clinical patterns of patients in our region. this is a descriptive retrospective study of 406 infertility cases presented at our urology clinic from February 2018 to February 2019. We assessed hormone and physical examination data, semen analysis results, the contribution of male and female factors to infertility, and types of infertility (i.e., primary or secondary). the age of the male patients ranged from 18 to 50 years, with a mean of 30 ± 5 years. Asthenozoospermia was the leading cause of male factor infertility in 77 patients (19%). Male factors as the sole cause of infertility were found in 185 (45.6%) couples. Female factors as the sole cause were found in 32 couples (7.9%). Primary infertility was determined in 314 (77.3%) patients, and 92 (22.7%) had secondary infertility. according to our results, the male infertility rate was high among couples reporting infertility. Couples should be informed about the causes of infertility, which may be due factors attributed to either sex.


Car backlight position and fog density bias observer-car distance estimates and time-to-collision judgments.
A series of experiments assessed biases in perceived distance that occur while driving as a function of the backlight position of the car ahead and fog density. V. Cavallo, M. Colomb, and J. Doré (2001) have shown that smaller horizontal backlight separation and fog may result in increased estimates of the distance between an observer and a car of which only the backlights are visible. They also predicted that raising the height of the car backlights would lead to increasing distance estimates. Distance perception was assessed in both static and dynamic computer-simulated scenarios in which the distance estimates were performed using a familiarized analog scale or using time-to-collision judgments for both pairs of backlights and single backlights. In a series of five experiments, the horizontal separation and fog density effects were replicated. In addition, distance estimates were consistently larger with higher than with lower vertical backlight positions. There is reason to believe that biases in distance perception may be augmented by car backlight positions and by low-visibility weather conditions. Car designers should take backlight placement seriously. Speed-dependent car-to-car distance control systems seem desirable to counteract biases in distance perception.


A systematic review of St. John's wort for major depressive disorder.
This systematic review evaluated St. John's wort (SJW) for the treatment of Major Depressive Disorder (MDD). The objectives of this review are to (1) evaluate the efficacy and safety of SJW in adults with MDD compared to placebo and active comparator and (2) evaluate whether the effects vary by severity of MDD. We searched PubMed, CINAHL, PsycINFO, CENTRAL, Embase, AMED, MANTIS, Web of Science, and ICTRP and existing reviews to November 2014. Two independent reviewers screened the citations, abstracted the data, and assessed the risk of bias. We included randomized controlled trials (RCTs) examining the effect of at least a 4-week administration of SJW on depression outcomes against placebo or active comparator in adults with MDD. Risk of bias was assessed using the Cochrane Risk of Bias tool and USPSTF criteria. Quality of evidence (QoE) was assessed using the GRADE approach. Thirty-five studies examining 6993 patients met inclusion criteria; eight studies evaluated a hypericum extract that combined 0.3 % hypericin and 1-4 % hyperforin. The herb SJW was associated with more treatment responders than placebo (relative risk [RR] 1.53; 95 % confidence interval [CI] 1.19, 1.97; I(2) 79 %; 18 RCTs; N = 2922, moderate QoE; standardized mean differences [SMD] 0.49; CI 0.23, 0.74; 16 RCTs; I(2) 89 %, N = 2888, moderate QoE). Compared to antidepressants, SJW participants were less likely to experience adverse events (OR 0.67; CI 0.56, 0.81; 11 RCTs; moderate QoE) with no difference in treatment effectiveness (RR 1.01; CI 0.90, 1.14; 17 RCTs, I(2) 52 %, moderate QoE; SMD -0.03; CI -0.21, 0.15; 14 RCTs; I(2) 74 %; N = 2248, moderate QoE) in mild and moderate depression. SJW monotherapy for mild and moderate depression is superior to placebo in improving depression symptoms and not significantly different from antidepressant medication. However, evidence of heterogeneity and a lack of research on severe depression reduce the quality of the evidence. Adverse events reported in RCTs were comparable to placebo and fewer compared with antidepressants. However, assessments were limited due to poor reporting of adverse events and studies were not designed to assess rare events. Consequently, the findings should be interpreted with caution. PROSPERO CRD42015016406 .


Cellular plasticity in reprogramming, rejuvenation and tumorigenesis: a pioneer TF perspective.
The multistep process of in vivo reprogramming, mediated by the transcription factors (TFs) Oct4, Sox2, Klf4, and c-Myc (OSKM), holds great promise for the development of rejuvenating and regenerative strategies. However, most of the approaches developed so far are accompanied by a persistent risk of tumorigenicity. Here, we review the groundbreaking effects of in vivo reprogramming with a particular focus on rejuvenation and regeneration. We discuss how the activity of pioneer TFs generates cellular plasticity that may be critical for inducing not only reprogramming and regeneration, but also cancer initiation. Finally, we highlight how a better understanding of the uncoupled control of cellular identity, plasticity, and aging during reprogramming might pave the way to the development of rejuvenating/regenerating strategies in a nontumorigenic manner.


Degradation of ornithine decarboxylase by the 26S proteasome.
Ornithine decarboxylase (ODC) is a key enzyme in polyamine biosynthesis. Turnover of ODC is extremely rapid and highly regulated, and is accelerated when polyamine levels increase. Polyamine-stimulated ODC degradation is mediated by association with antizyme (AZ), an ODC inhibitory protein induced by polyamines. ODC, in association with AZ, is degraded by the 26S proteasome in an ATP-dependent, but ubiquitin-independent, manner. The 26S proteasome irreversibly inactivates ODC prior to its degradation. The inactivation, possibly due to unfolding, is coupled to sequestration of ODC within the 26S proteasome. This process requires AZ and ATP, but not proteolytic activity of the 26S proteasome. The carboxyl-terminal region of ODC presumably exposed by interaction with AZ plays a critical role for being trapped by the 26S proteasome. Thus, the degradation pathway of ODC proceeds as a sequence of multiple distinct processes, including recognition, sequestration, unfolding, translocation, and ultimate degradation mediated by the 26S proteasome.


Capsule Machine Validation.
Instrumentation has become an integral function for any compounding operation, particularly for hard-gelatin capsules. This study examined the use of a capsule machine in a simulated compounding operation. It is an instrument that is reasonably priced and can deliver 24 capsules within ten minutes of operation. Capsule Connection LLC suggests the use of this machine for filling capsules with herbal or vitamin preparations. Moreover, this instrument may be used for training pharmacy students in a pharmaceutics/compounding laboratory. The main objective of the research was to calibrate and validate a capsule formulation for acetaminophen using the Capsule Machine from Capsule Connection LLC. Twenty Capsule Machines were purchased from Capsule Connection LLC. The machines were first calibrated and then validated using spray-dried lactose monohydrate powder. Based on the results obtained from the calibration and validation process, a powder mixture containing acetaminophen was prepared by mixing acetaminophen with lactose in a glass mortar using the geometric dilution method. The labeled amount of acetaminophen per capsule was 81 mg. Capsules generated from this compounding operation were tested using the United States Pharmacopeia content uniformity test. The calibration and validation processes were repeated twice with lactose monohydrate. The overall average weights for size-0 capsule shells and lactose-filled capsules were 94.61 mg and 545.77 mg, respectively. On average, the content of a capsule was estimated to be 451.16 mg of powder. A powder mixture was prepared to make 480 capsules (20 machines, each producing 24 capsules). This mixture was made of 38.88 g of acetaminophen and 177.68 g of lactose. A 3% amount of additional powder was added to account for the powder loss during the operation. All capsule machines passed the United States Pharmacopeia content uniformity test. The average amount of acetaminophen per capsule was 98.91% of the labeled amount. Only less than 5% of the capsules were lost during the filling operation of lactose in capsules and none lost with the acetaminophen capsules. The Capsule Machine from Capsule Connection LLC produced capsules containing acetaminophen with high reproducibility and accuracy.


Dual-Energy Computed Tomography Imaging in Early-Stage Hepatocellular Carcinoma: A Preliminary Study.
This study aims to evaluate the application of dual-energy computed tomography (DECT) for multiparameter quantitative measurement in early-stage hepatocellular carcinoma (HCC). The study retrospectively enrolled 30 patients with early-stage HCC and 43 patients with early-stage HCC who received radiofrequency ablation (RFA) and underwent abdomen enhanced CT scans in GSI mode. The GSI viewer was used for image display and data analysis. The regions of interest (ROIs) were delineated in the arterial phase and the venous phase. The optimal single energy value, CT values on different energy levels (40 keV, 70 keV, 100 keV, and 140 keV), the optimal energy level, the slope of the spectral attenuation curve, the effective atomic number (Z eff), iodine concentration (IC), water concentration (WC), normalized iodine concentration (NIC), and normalized water concentration (NWC) are measured and quantitatively analyzed. The CT values of early-stage HCC at different single energy levels in dual phases were significantly different, and the single energy values were negatively correlated with the CT values. In the arterial phase and the venous phase, the optimal energy values for the best contrast-to-noise ratio were (68.34 ± 3.20) keV and (70.14 ± 2.01) keV, respectively. The slope of the spectral attenuation curve showed a downward trend at 40 keV, 70 keV, 100 keV, and 140 keV, but there was no statistically significant difference (P > 0.05). Z eff was positively correlated with IC and standardized IC, but has no significant correlation with WC and NWC in dual phases. DECT imaging contains multiparameter information and has different application values for early-stage HCC, and it is necessary to select the parameters reasonably for personalized and comprehensive evaluation.


Associations between sleep disorders, sleep duration, quality of sleep, and hypertension: results from the National Health and Nutrition Examination Survey, 2005 to 2008.
Sleep is a contributing factor to optimal health and vitality. However, to date, no national study has evaluated the simultaneous relationship between sleep disorders, quality, and duration with hypertension. Using data from National Health and Nutrition Examination Survey (NHANES) (2005 to 2008), hypertension was defined by current use of antihypertensive medication or systolic blood pressure ≥140 mm Hg or diastolic blood pressure ≥90 mm Hg. Self-reported sleep disorders and duration were categorized from a single household interview question, and sleep quality was determined from several questions on sleeping habits. The prevalence of hypertension was 30.2% and 7.5%, and 33.0% and 52.1% reported having sleep disorders, short sleep, and poor sleep, respectively. After adjustment for demographic characteristics and comorbidities, having sleep disorders only was not significantly associated with hypertension (odds ratio [OR], 1.65; 95% confidence interval [CI], 0.73-3.77). However, this association was modified by sleep duration: significant associations were observed among adults with concurrent sleep disorders and short sleep (OR, 2.30; 95% CI, 1.49-3.56) and with sleep disorders, short sleep, and poor sleep (OR, 1.84; 95% CI, 1.13-2.98). These findings indicate an association between a combination of sleep problems and hypertension, but prospective studies are needed to understand the complex interplay between them.


Renal medullary microcirculation: architecture and exchange.
The architecture of the renal medullary microcirculation is highly specialized. Consistent with their role in countercurrent exchange, the vessels (the vasa recta and the intervening capillaries) have high permeabilities to fluid and small hydrophilic solutes. The urea permeability of the continuous endothelium of the descending vasa recta (DVR) in the outer medulla is greatly enhanced by a urea transporter. Aquaporin channels have also been identified in these vessels. In spite of the absence of lymphatics from the inner medulla, fluid uptake from the interstitial fluid (ISF) through the fenestrated endothelium of the ascending vasa recta (AVR) appears to be driven by differences in hydrostatic and oncotic pressure. Because the AVR have high Lp's [10(-5) cm s-1 (cm H2O)-1] and are mechanically linked to surrounding structures, small increments of ISF pressures above the pressure within the AVR can drive significant volumes of fluid into AVR if ISF volume expands. The lower reflection coefficients to serum albumin of the AVR as compared with the DVR may be important in the clearance of interstitial plasma protein. Recent work on isolated DVR from the outer medulla has revealed that these vessels are capable of vasoconstriction and thus of regulating medullary blood flow.


A new expandable cannula system for endoscopic evacuation of intraparenchymal hemorrhages.
The authors describe a newly developed expandable cannula to enable a more efficient use of an endoscope in removing intraparenchymal spontaneous hypertensive intracerebral hematomas. The cannula is introduced like a conventional brain cannula, using neuronavigation techniques to reach the targeted hematoma accurately, and, once deployed, conventional microsurgical techniques are used under direct endoscopic visualization. This method was used in 6 patients, and, based on the results of intraoperative intracranial pressure monitoring and postoperative CT scanning, the authors were able to achieve good hematoma removal. They found that by using the expandable cannula, efficient endoscopic surgery in the brain parenchyma was possible.


Raynaud phenomenon in children.
Question I have several patients, mostly girls, who are living with Raynaud phenomenon. Does this condition appear in children, and what should be the course of action?Answer Raynaud phenomenon, described in the 1860s, can present in children and even in the first decade of life. While most children will have primary Raynaud phenomenon, with no serious adverse consequences, in others it might be a sign of a pending systemic disease. Those children with a positive reaction to antinuclear antibody, specific autoantibodies associated with connective tissue disease, or nail fold capillary changes require referral to a pediatric rheumatologist and close follow-up.


Residue iteration decomposition (RIDE): A new method to separate ERP components on the basis of latency variability in single trials.
Event-related brain potentials (ERPs) are important research tools because they provide insights into mental processing at high temporal resolution. Their usefulness, however, is limited by the need to average over a large number of trials, sacrificing information about the trial-by-trial variability of latencies or amplitudes of specific ERP components. Here we propose a novel method based on an iteration strategy of the residues of averaged ERPs (RIDE) to separate latency-variable component clusters. The separated component clusters can then serve as templates to estimate latencies in single trials with high precision. By applying RIDE to data from a face-priming experiment, we separate priming effects and show that they are robust against latency shifts and within-condition variability. RIDE is useful for a variety of data sets that show different degrees of variability and temporal overlap between ERP components.


Disparate intracellular processing of human IL-12 preprotein subunits: atypical processing of the P35 signal peptide.
IL-12 is a heterodimeric cytokine produced by APC that critically regulates cell-mediated immunity. Because of its crucial function during immune responses, IL-12 production is stringently regulated, in part through transcriptional control of its p35 subunit, which requires the differentiative effects of IFN-gamma for expression. To determine whether post-transcriptional aspects of IL-12 production might be regulated, we examined intracellular protein processing of each subunit. We report here that p40 and p35 subunits are processed by disparate pathways. Whereas processing of p40 conforms to the cotranslational model of signal peptide removal concomitant with translocation into the endoplasmic reticulum (ER), processing of p35 does not. Translocation of the p35 preprotein into the ER was not accompanied by cleavage of the signal peptide; rather, removal of the p35 signal peptide occurred via two sequential cleavages. The first cleavage took place within the ER, and the cleavage site localized to the middle of the hydrophobic region of the signal peptide. Although the preprotein was glycosylated upon entry into the ER, its glycosylation status did not affect primary cleavage. Subsequently, the remaining portion of the p35 signal peptide was removed by a second cleavage, possibly involving a metalloprotease, concomitant with additional glycosylation and secretion. Secretion could be inhibited by mutation of the second cleavage site or by inhibition of glycosylation with tunicamycin. In contrast, p40 secretion was not affected by inhibition of glycosylation. Our findings demonstrate that IL-12 subunits are processed by disparate pathways and suggest new modalities for regulation of IL-12 production.


Piezoelectric effects and electromechanical theories at the nanoscale.
Considerable effort has been made to study the piezoelectric effect on the nanoscale, which serves as a physical basis for a wide range of smart nanodevices and nanoelectronics. This paper reviews recent progress in the research on the piezoelectric properties and electromechanical effects of piezoelectric nanomaterials (PNs). The review begins with an introduction to existing PNs which exhibit a diverse range of atomic structures and configurations. The nanoscale measurement of their effective piezoelectric coefficients (EPCs) is summarised with an emphasis on the major factors determining the piezoelectric properties of PNs. The paper concludes with a review of the electromechanical theories that are able to capture the small-scale effects on PNs, which include the surface piezoelectricity, flexoelectricity and Eringen's nonlocal theory. In contrast to the classical theories, two types of EPCs are defined, which were found to be size-dependent and loading condition-selective.


Italian risk factor-based screening for gestational diabetes.
There is a debate about whether universal or risk factors-based screening is most appropriate for gestational diabetes diagnosis. The aim of our retrospective study was to compare in our population the universal screening test recommended by the International Association of Diabetes in Pregnancy Study Group (IADPSG) panel and the American Diabetes Association (ADA) versus the selective screening proposed by the United Kingdom National Institute for Health and Clinical Excellence guidelines (NICE) but modified by the Italian National Institute of Health. From May 2010 to October 2011 all consecutive pregnant women were screened for gestational diabetes according to the IADPSG's panel criteria, while all the risk factors for each patient were registered. Of the 1015 pregnant women included in the study, 113 (11%) were diagnosed with gestational diabetes and 26 (23%) of them would not have been identified by the selective screening proposed by the Italian National Institute of Health. However, all the risk factors considered by the selective screening revealed a good predictive role except for maternal age ≥ 35 years (OR: 0.98). In the group without the risk factors considered, it was reported the predictive role for gestational diabetes of prepregnancy BMI and nulliparity. The selective risk factors-based screening proposed by the Italian National Institute of Health has detected 77% of gestational diabetes cases in our population, sparing the oral glucose tolerance test for more than 40% of pregnant women at the same time. More information on the clinical impact of this choice could be obtained by a strict analysis of treatment, perinatal outcome and follow-up of an adequate sample size of "missed" gestational diabetes.


Neural regulation of alpha-dystroglycan biosynthesis and glycosylation in skeletal muscle.
Alpha-dystroglycan (alpha-DG) is part of a complex of cell surface proteins linked to dystrophin or utrophin, which is distributed over the myofiber surface and is concentrated at neuromuscular junctions. In laminin overlays of muscle extracts from developing chick hindlimb muscle, alpha-DG first appears at embryonic day (E) 10 with an apparent molecular mass of 120 kDa. By E15 it is replaced by smaller (approximately 100 kDa) and larger (approximately 150 kDa) isoforms. The larger form increases in amount and in molecular mass (>200 kDa) as the muscle is innervated and the postsynaptic membrane differentiates (E10-E20), and then decreases dramatically in amount after hatching. In myoblasts differentiating in culture the molecular mass of alpha-DG is not significantly increased by their replication, fusion, or differentiation into myotubes. Monoclonal antibody IIH6, which recognizes a carbohydrate epitope on alpha-DG, preferentially binds to the larger forms, suggesting that the core protein is differentially glycosylated beginning at E16. Consistent with prior observations implicating the IIH6 epitope in laminin binding, the smaller forms of alpha-DG bind more weakly to laminin affinity columns than the larger ones. In blots of adult rat skeletal muscle probed with radiolabeled laminin or monoclonal antibody IIH6, alpha-DG appears as a >200-kDa band that decreases in molecular mass but increases in intensity following denervation. Northern blots reveal a single mRNA transcript, indicating that the reduction in molecular mass of alpha-DG after denervation is not obviously a result of alternative splicing but is likely due to posttranslational modification of newly synthesized molecules. The regulation of alpha-DG by the nerve and its increased affinity for laminin suggest that glycosylation of this protein may be important in myofiber-basement membrane interactions during development and after denervation.


Removal of arsenic from water using the adsorbent: New Zealand iron-sand.
Adsorption is a technology used to remove arsenic from water contaminated at levels above drinking water standards. In this study, New Zealand Iron-Sand (NZIS), a naturally-available adsorbent was investigated for its efficiency in removing both As (III) and As (V). Several batch tests were conducted with different concentrations of arsenic at different pH conditions. During the batch tests, the maximum adsorption of As (III) occurred at a pH of 7.5, while As (V) adsorption reached its maximum value at a pH of 3. Both Langmuir and Freundlich adsorption models were found to fit with R(2) values greater than 0.92. From the Langmuir adsorption model, the maximum adsorption capacity of NZIS for As (III) and As (V) were estimated to be 1,250 and 500 μg/g, respectively. These values were substantial enough to consider NZIS a promising new adsorbent for arsenic removal.


Aberrant promoter methylation of EDNRB in lung cancer in Taiwan.
Hypermethylation has been shown in the promoter region of the endothelin receptor B (EDNRB) gene in several human tumors, but its role in lung cancer formation is unclear. In this study, genomic DNA from lung cancer patients was subjected to methylation-specific PCR to determine the methylation status of the EDNRB gene in lung cancer. Aberrant methylation of the EDNRB gene was detected in 32.9% (26 of 79) lung cancer patients. Promoter hypermethylation of EDNRB was found to significantly differ with histological type but was not correlated to other clinicopathological characteristics. Decreased mRNA transcripts were correlated to aberrant methylation. Treatment with 5-aza-deoxycytidine reversed the methylation status and re-expression of the EDNRB gene in the H1355 human lung cancer cell line. Our results suggest that inactivation of the EDNRB gene through epigenetic alteration is highly prevalent in lung cancer in Taiwan.


Juvenile rhabdomyosarcomas in two dogs.
Juvenile rhabdomyosarcomas were diagnosed in two young dogs based on the results of histopathology, phosphotungstic acid-hematoxylin stain, immunohistochemistry, and the age of the dogs. One dog, an 11-month-old Rottweiler, had tumor masses in the maxillary gingiva and the urinary bladder. Histologically, the gingival mass was an alveolar type of rhabdomyosarcoma and the urinary bladder mass was an embryonal type. The other dog, a 1.5-year-old Basset Hound, had an embryonal rhabdomyosarcoma involving the oropharynx and the temporal muscles, with metastasis to the regional lymph node and lungs.