Flat flexible school shoes for adolescents with patellofemoral pain: a randomised, assessor-blinded, parallel-group feasibility trial.
To determine the feasibility of conducting a large-scale randomised controlled trial on the efficacy of flat, flexible school footwear versus traditional school footwear in adolescents (aged 12-18 years) with patellofemoral pain (PFP). Adolescents with PFP were recruited for this study. Participants were randomised to wear either a (1) flat, flexible school shoe or (2) a traditional school shoe. Participants wore the shoes as per school requirements for 12 weeks. Feasibility was assessed by (1) adherence to allocated shoe wear of ≥75% of total weekly school shoe wear time (recorded through weekly log sheets), (2) a recruitment rate of one participant per fortnight and (3) a dropout rate of ≤ 20%. Descriptive statistics were used for feasibility outcomes. 24 adolescents (15 men, 9 women, mean (SD) age 14.3 (1.7) years) participated in this study. Two participants (8%) were lost to follow-up. The recruitment rate was 1.7 participants per fortnight. 11 of 12 participants (91%) in the flat flexible shoe group and 9 of 10 participants (90%) in the traditional shoe group met the minimum adherence for shoe wear. Mean weekly shoe wear was 20 (7.6) and 21 (4.5) hours per week in the flat, flexible, and traditional shoe groups, respectively. Our results indicate that progression to a full-scale randomised controlled trial is feasible based on the current protocol. A full-scale randomised controlled trial powered to detect estimates of treatment efficacy using flat, flexible school shoes versus traditional school shoes is warranted and will guide evidence-based management of adolescent PFP.


Development of the human brain: in vivo quantification of metabolite and water content with proton magnetic resonance spectroscopy.
Cerebral metabolite concentrations and water content were measured by means of localized proton magnetic resonance spectroscopy in 50 children, while metabolite peak ratios in short echo time spectra were evaluated in 173 examinations. Normative curves for normal development were established for two cerebral locations. The current report presents the first study of absolute metabolite concentrations and T1- and T2- relaxation as a function of age. Myo-inositol was found dominating the spectra at birth (12 mmoles/kg), while choline is responsible for the strongest peak in older infants (2.5 mmoles/kg). Creatine and N-acetyl groups are at significantly lower concentrations in the neonate than in the adult (Cr: 6, NA: 5 mmoles/kg). NA and Cr are determined by gestational age, whereas the concentration of ml correlates best with postnatal age. Quantitative 1H MRS is expected to be of particular value in diagnosis and monitoring of pathology in infants, since metabolite ratios are often misleading.


Abnormal tibiofemoral kinematics following ACL reconstruction are associated with early cartilage matrix degeneration measured by MRI T1rho.
Altered kinematics following ACL-reconstruction may be a cause of post-traumatic osteoarthritis. T(1ρ) MRI is a technique that detects early cartilage matrix degeneration. Our study aimed to evaluate kinematics following ACL-reconstruction, cartilage health (using T(1ρ) MRI), and assess whether altered kinematics following ACL-reconstruction are associated with early cartilage degeneration. Eleven patients (average age: 33 ± 9 years) underwent 3T MRI 18 ± 5 months following ACL-reconstruction. Images were obtained at extension and 30° flexion under simulated loading (125 N). Tibial rotation (TR) and anterior tibial translation (ATT) between flexion and extension, and T(1ρ) relaxation times of the knee cartilage were analyzed. Cartilage was divided into five compartments: medial and lateral femoral condyles (MFC/LFC), medial and lateral tibias (MT/LT), and patella. A sub-analysis of the femoral weight-bearing (wb) regions was also performed. Patients were categorized as having "abnormal" or "restored" ATT and TR, and T(1ρ) percentage increase was compared between these two groups of patients. As a group, there were no significant differences between ACL-reconstructed and contralateral knee kinematics, however, there were individual variations. T(1ρ) relaxation times of the MFC and MFC-wb region were elevated (p ≤ 0.05) in the ACL-reconstructed knees compared to the uninjured contralateral knees. There were increases (p ≤ 0.05) in the MFC-wb, MT, patella and overall average cartilage T(1ρ) values of the "abnormal" ATT group compared to "restored" ATT group. The percentage increase in the T(1ρ) relaxation time in the MFC-wb cartilage approached significance (p=0.08) in the "abnormal" versus "restored" TR patients. Abnormal kinematics following ACL-reconstruction appear to lead to cartilage degeneration, particularly in the medial compartment.


Calmodulin antagonizes amyloid-β peptides-mediated inhibition of brain plasma membrane Ca(2+)-ATPase.
The synaptosomal plasma membrane Ca(2+)-ATPase (PMCA) plays an essential role in regulating intracellular Ca(2+) concentration in brain. We have recently found that PMCA is the only Ca(2+) pump in brain which is inhibited by amyloid-β peptide (Aβ), a neurotoxic peptide implicated in the pathology of Alzheimer's disease (AD) [1], but the mechanism of inhibition is lacking. In the present study we have characterized the inhibition of PMCA by Aβ. Results from kinetic assays indicate that Aβ aggregates are more potent inhibitors of PMCA activity than monomers. The inhibitory effect of Aβ could be blocked by pretreating the purified protein with Ca(2+)-calmodulin, the main endogenous activator of PMCA, and the activity of truncated PMCA lacking the calmodulin binding domain was not affected by Aβ. Dot-overlay experiments indicated a physical association of Aβ with PMCA and also with calmodulin. Thus, calmodulin could protect PMCA from inhibition by Aβ by burying exposed sites on PMCA, making them inaccessible to Aβ, and also by direct binding to the peptide. These results suggest a protective role of calmodulin against neuronal Ca(2+) dysregulation by PMCA inhibition induced by Aβ.


Mixed Matrix Membranes for Efficient CO2 Separation Using an Engineered UiO-66 MOF in a Pebax Polymer.
Mixed matrix membranes (MMMs) have attracted significant attention for overcoming the limitations of traditional polymeric membranes for gas separation through the improvement of both permeability and selectivity. However, the development of defect-free MMMs remains challenging due to the poor compatibility of the metal-organic framework (MOF) with the polymer matrix. Thus, we report a surface-modification strategy for a MOF through grafting of a polymer with intrinsic microporosity onto the surface of UiO-66-NH2. This method allows us to engineer the MOF-polymer interface in the MMMs using Pebax as a support. The insertion of a PIM structure onto the surface of UiO-66-NH2 provides additional molecular transport channels and enhances the CO2 transport by increasing the compatibility between the polymer and fillers for efficient gas separation. As a result, MMM with 1 wt% loading of PIM-grafted-MOF (PIM-g-MOF) exhibited very promising separation performance, with CO2 permeability of 247 Barrer and CO2/N2 selectivity of 56.1, which lies on the 2008 Robeson upper bound. Moreover, this MMM has excellent anti-aging properties for up to 240 days and improved mechanical properties (yield stress of 16.08 MPa, Young's modulus of 1.61 GPa, and 596.5% elongation at break).


An Anti-microbial Terpenoid Fraction from Gymnema sylvestre Induces Flip-flop of Fluorescent-Phospholipid Analogs in Model Membrane.
Therapeutic potential of Gymnema sylvestre on diverse cell types is predominantly due to a variety of terpenoids and their derivatives. However, their bioavailability becomes limited due to poor solubility and lower lipophilic properties, provoking the search for novel membranotropic terpenoids and their mechanism of action. A terpenoid fraction purified from Gymnema sylvestre exhibited broad spectrum antimicrobial activity against both Gram positive and Gram negative bacteria with IC50 ˂ 0.1 mg/ml. Evaluation of its membranotropic effect in vitro on reconstituted model membrane revealed that the fraction induced flip-flop of fluorescent phospholipid analogs across the lipid bilayer. The terpenoid-induced lipid flipping was biphasic with a fast linear phase (rate constant (k1) = 3 to 5 S-1) and a second slow exponential phase (rate constant (k2) = (4 to 9) × 10-3 S-1). The lipid-flippase activity of the terpenoid fraction showed concentration and incubation-dependent cooperativity, indicating their lipophilic nature and membrane-destabilizing activity that facilitated lipid translocation. For the first time, our study reveals the flippase activity of a terpenoid fraction of Gymnema sylvestre that could be further explored for their membrane-mediated pharmacological properties. Graphical Abstract.


Acetaminophen does not impair clearance of zidovudine.
To determine whether concurrent treatment with acetaminophen and zidovudine impairs clearance of zidovudine, thereby increasing the risk for zidovudine-induced hematologic toxicity. Dose escalation, drug interaction study. University clinical research center. Patients with the acquired immunodeficiency syndrome (AIDS) or advanced AIDS-related complex. Acetaminophen and 200 mg of zidovudine simultaneously every 4 hours. For 13 patients, the unit dosage of acetaminophen was 325 mg for 3 days; for 8 patients, the dosage was 650 mg for 3 days; and, for 6 patients, the dosage was 650 mg for 7 days. Zidovudine clearance and production of the glucuronide conjugate of zidovudine were assessed after acetaminophen treatment. Neither zidovudine clearance nor production of the glucuronide conjugate of zidovudine was impaired after treatment with acetaminophen. Clearance of zidovudine was actually accelerated by 5%, 11%, and 33% with the three acetaminophen regimens, respectively (P = 0.002 by analysis of variance; P = 0.04 for linear trend when changes in the area-under-the-curve for zidovudine were compared). Because serum concentrations of zidovudine decrease after the coadministration of acetaminophen, a pharmacokinetic interaction between zidovudine and acetaminophen is unlikely to increase the risk for hematologic toxicity associated with zidovudine.


Characterization of lncRNA functioning in ovine conceptuses and endometria during the peri-implantation period.
In ruminants, RNA-sequence analyses have revealed many characteristics of transcripts expressed in conceptuses (embryo and extraembryonic membrane) during peri-implantation periods; however, lncRNA profiles are yet characterized. In this study, we aimed to characterize the lncRNA expression profile in conceptuses during peri-implantation periods in sheep. We analyzed the RNA-sequence data of ovine conceptuses and endometria obtained from pregnant animals on days 15, 17, 19 and 21 (day 0 = day of estrus, n = 3 or 4/day). We predicted the protein coding ability of the assembled transcripts to identify the lncRNA candidates. This analysis identified 8808 lncRNAs, 3423 of which were novel lncRNAs. Gene ontology analysis revealed that lncRNA target genes were enriched for biological processes involved in the respiratory electron transport chain (RETC). qPCR analysis demonstrated that the expression levels on transcripts encoding RETC such as mitochondrially encoded cytochrome c oxidase II (MTCO2) and mitochondria DNA copy number in conceptuses were not increased on P21, although western blotting analysis and immunohistochemistry demonstrated that MTCO2 protein in conceptuses was increased on P21. NAD/NADH assay revealed that NADH level in conceptuses was increased on P21. These results indicate that lncRNAs could regulate the RETC through post-transcriptional levels in the conceptuses. Therefore, lncRNA is a potential new regulator in ovine conceptus development during peri-implantation periods.


Should individuals be informed about their salt sensitivity status? First indications of the value of testing for genetic predisposition to low-risk conditions.
The present study examined the possible pathways of effect of genetic testing for relatively "low-risk" conditions by exploring positive as well as negative effects of anticipated test results on the intention to restrict salt intake. In a cross-sectional within-subjects design, patients being tested for genetic predispositions to salt sensitivity reported higher overall intentions to restrict their salt intake when anticipating positive test results, confirming the value of genetic testing for low-risk conditions. However, participants in the precontemplation and preparation stages of change reported lower intentions when anticipating negative test results. This result suggests that negative test results have a negative impact on the motivation to perform preventive behavior among those who have not yet considered, and those who are planning to perform, the preventive behavior. Although the results show that, overall, genetic testing for low-risk conditions has a positive impact on the motivation to engage in preventive behavior, one needs to be aware of the potential negative effect of receiving negative test results. Consequently, individuals receiving negative test results should be carefully counseled on the meaning of the results and the consequences for preventive behavior.


Composite biomarkers for assessing Duchenne muscular dystrophy: an initial assessment.
Compared with individual parameters, composite biomarkers may provide a more effective means for monitoring disease progression and the effects of therapy in clinical trials than single measures. In this study, we built composite biomarkers for use in Duchenne muscular dystrophy by combining values from two objective measures of disease severity: electrical impedance myography and quantitative ultrasound and evaluating how well they correlated to standard functional measures. Using data from an ongoing study of electrical impedance myography and quantitative ultrasound in 31 Duchenne muscular dystrophy and 26 healthy boys aged 2-14 years, we combined data sets by first creating z scores based on the normal subject data and then using simple mathematical operations (addition and multiplication) to create composite measures. These composite scores were then correlated to age and standard measures of function including the 6-minute walk test, the North Star Ambulatory Assessment, and handheld dynamometry. Combining data sets resulted in stronger correlations with all four outcomes than for either electrical impedance myography or quantitative ultrasound alone in six of eight instances. These improvements reached statistical significance (P < 0.05) in several cases. For example, the correlation coefficient for the composite measure with the North Star Ambulatory Assessment was 0.79 but was only 0.66 and 0.67 (respectively) for gray scale level and electrical impedance myography separately. Arithmetically derived composite scores can provide stronger correlations to functional measures than isolated biomarkers. Longitudinal study of such composite markers in Duchenne muscular dystrophy clinical trials is warranted.


Photonic Hyperthermia Synergizes with Immune-Activators to Augment Tumor-Localized Immunotherapy.
Photothermal immunotherapy, the combination of photothermal hyperthermia and immunotherapy, is a noninvasive and desirable therapeutic strategy to address the deficiency of traditional photothermal ablation for tumor treatment. However, insufficient T-cell activation following photothermal treatment is a bottleneck to achieve satisfactory therapeutic effectiveness. In this work, a multifunctional nanoplatform is rationally designed and engineered on the basis of polypyrrole-based magnetic nanomedicine modified by T-cell activators of anti-CD3 and anti-CD28 monoclonal antibodies, which have achieved robust near infrared laser-triggered photothermal ablation and long-lasting T-cell activation, realizing diagnostic imaging-guided immunosuppressive tumor microenvironment regulation following photothermal hyperthermia by reinvigorating tumor-infiltrating lymphocytes. By virtue of high-efficient immunogenic cell death and dendritic cell maturation combined with T-cell activation, this nanosystem markedly restrains primary and abscopal tumors as well as metastatic tumors with negligible side effects in vivo, exerting the specific function for suppressing tumor recurrence and metastasis by establishing a long-term memory immune response.


Adulteration and Contamination of Commercial Sap of Hymenaea Species.
The Hymenaea stigonocarpa and Hymenaea martiana species, commonly known as "jatobá," produce a sap which is extracted by perforation of the trunk and is commonly used in folk medicine as a tonic. For this study, the authenticity of commercial samples of jatobá was verified by the identification of the main compounds and multivariate analysis and contamination by microbial presence analysis. The acute toxicity of the authentic jatobá sap was also evaluated. The metabolites composition and multivariate analysis revealed that none of the commercial samples were authentic. In the microbiological contamination analysis, five of the six commercial samples showed positive cultures within the range of 1,700-100,000 CFU/mL and the authentic sap produced no signs of toxicity, and from a histological point of view, there was the maintenance of tissue integrity. In brief, the commercial samples were deemed inappropriate for consumption and represent a danger to the population.


A Quieter Ocean: Experimentally Derived Differences in Attentive Responses of Tursiops truncatus to Anthropogenic Noise Playbacks before and during the COVID-19-Related Anthropause.
The effects of anthropogenic noise continue to threaten marine fauna, yet the impacts of human-produced sound on the broad aspects of cognition in marine mammals remain relatively understudied. The shutdown of non-essential activities due to the COVID-19-related anthropause created an opportunity to determine if reducing levels of oceanic anthropogenic noise on cetaceans affected processes of sensitization and habituation for common human-made sounds in an experimental setting. Dolphins at Dolphin Quest Bermuda were presented with three noises related to human activities (cruise ship, personal watercraft, and Navy low-frequency active sonar) both in 2018 and again during the anthropause in 2021 via an underwater speaker. We found that decreased anthropogenic noise levels altered dolphin responses to noise playbacks. The dolphins spent significantly more time looking towards the playback source, but less time producing burst pulse and echolocation bouts in 2021. The dolphins looked towards the cruise ship sound source significantly more in 2021 than 2018. These data highlight that different sounds may incur different habituation and sensitization profiles and suggest that pauses in anthropogenic noise production may affect future responses to noise stimuli as dolphins dishabituate to sounds over time.


Non-canonical role of wild-type SEC23B in the cellular stress response pathway.
While germline recessive loss-of-function mutations in SEC23B in humans cause a rare form of anaemia, heterozygous change-of-function mutations result in increased predisposition to cancer. SEC23B encodes SEC23 homologue B, a component of coat protein complex II (COPII), which canonically transports proteins from the endoplasmic reticulum (ER) to the Golgi. Despite the association of SEC23B with anaemia and cancer, the precise pathophysiology of these phenotypic outcomes remains unknown. Recently, we reported that mutant SEC23B has non-canonical COPII-independent function, particularly within the ER stress and ribosome biogenesis pathways, and that may contribute to the pathobiology of cancer predisposition. In this study, we hypothesized that wild-type SEC23B has a baseline function within such cellular stress response pathways, with the mutant protein reflecting exaggerated effects. Here, we show that the wild-type SEC23B protein localizes to the nucleus in addition to classical distribution at the ER/Golgi interface and identify multiple putative nuclear localization and export signals regulating nuclear-cytoplasmic transport. Unexpectedly, we show that, independently of COPII, wild-type SEC23B can also localize to cell nucleoli under proteasome inhibition conditions, with distinct distribution patterns compared to mutant cells. Unbiased proteomic analyses through mass spectrometry further revealed that wild-type SEC23B interacts with a subset of nuclear proteins, in addition to central proteins in the ER stress, protein ubiquitination, and EIF2 signalling pathways. We validate the genotype-specific differential SEC23B-UBA52 (ribosomal protein RPL40) interaction. Finally, utilizing patient-derived lymphoblastoid cell lines harbouring either wild-type or mutant SEC23B, we show that SEC23B levels increase in response to ER stress, further corroborating its role as a cellular stress response sensor and/or effector. Overall, these observations suggest that SEC23B, irrespective of mutation status, has unexplored roles in the cellular stress response pathway, with implications relevant to cancer and beyond that, CDAII and normal cell biology.


Best Practices for Depression Screening Among College Students: A Literature Review.
Rates of depression among the college-aged population are alarming, particularly in light of the coronavirus disease 2019 pandemic. It is important for providers who care for this population to have proper screening and referral processes to meet growing student needs. The purpose of this literature review was to examine current best practices for depression screening among college students. Comprehensive searches of PubMed, CINAHL, Embase, and Cochrane databases were completed with the assistance of a medical librarian. A total of 775 studies were screened, and 17 articles were included in the review. Evidence supports universal-based depression screenings; use of screening methods to include brief and online screenings; use of validated screening tools, including the Patient Health Questionnaire (PHQ) -2 and PHQ-9; and the importance of a proper referral process and adequate access to treatment measures. An appropriate depression screening and referral process within campus health centers is critical to meet the needs of college students. [Journal of Psychosocial Nursing and Mental Health Services, xx(x), xx-xx.].


Temporal trends in pulmonary embolism prevalence in Greece during 2013-2017.
Pulmonary embolism (PE) epidemiological data about the disease prevalence in the general population are unclear. The present study aims to investigate the prevalence of PE in Greece and the associated temporal trends for the years 2013-2017. Data on medical prescriptions for PE in the years 2013-2017 were provided by the Greek National Health Service Organization (EOPYY). Data on age, gender, specialty of the prescribing physician and prescription unit were provided as well. The total number of medical prescriptions for PE for the study period was 101,426. Of the total prescriptions, 51% were issued by the Public Sector and 48% by the Private Sector. In 2013 the prevalence of PE was 5.43 cases per 100,000 citizens and increased constantly until 2017 with 23.79 cases per 100,000 population. Prevalence was higher in all years studied in the age group of 70-80 years. For the year 2017, we observed 69.35 cases per 100,000 population for subjects 70-80 years, followed by the ages 80-90 (60.58/100,000) and 60-70 years (56.47 /100,000). Females displayed higher PE prevalence than males and higher increasing trend. PE prevalence has an increasing trend throughout the years 2013-2017 while prevalence in females is higher than males and displays a higher increasing trend. Our results may be used to appropriately organize nationwide health care campaigns aiming at the diagnosis, treatment and prevention of PE.


Dyspnea: perfusion index and triage status.
To determine the relationship between perfusion index and the emergency triage classification in patients admitted to the emergency department with dyspnea. Adult patients who presented with dyspnea and whose perfusion index values were measured with Masimo Radical-7 device at the time of admission, at the first hour and the second hour of admission were included in the study. The PI and oxygen saturation measured by finger probes were compared and the superiority of their effects on the emergency triage classification was compared. For the 0.9 cut- off value of the arrival PI level according to the triage status; sensitivity 79.25%; specificity 78.12%; positive predictive value is 66.7 and negative predictive value is 87.2. A statistically significant correlation was found between the triage status and the 0.9 cut- off value of the admission PI level. We can say that the ODDS rate of red triage is 13.63 times (95% CI: 5.99-31.01) times higher in cases with a PI level of 0.9 and below. In the ROC analysis, the cut-off value of 1.1 and above the admission PI level was determined as the most appropriate point for discharge. The perfusion index can help to determine the triage classification in emergency departments for dyspnea.


[Distribution of enterobacteria in man and small mammals].
The species composition of the most common enterobacteria isolated from the intestine of 825 patients with intestinal dysbacteriosis and 292 small mammals of different species is compared.


The impact of the menstrual cycle on intra-operative and postoperative bleeding in abdominoplasty patients.
Hormonal changes that take place during the menstrual cycle may have an effect on perioperative bleeding. Previous studies have confirmed a relationship between the menstrual cycle and surgical bleeding in rhinoplasty and breast reduction surgery; yet both surgical sites display changes in accordance with sex hormone levels during the menstrual phase. The aim of this study was to determine the effect of the menstrual cycle on perioperative bleeding when undergoing surgeries of sites not directly related to menstrual hormonal changes. Forty-one patients undergoing abdominoplasty were grouped as group A - perimenstrual (0-7, 21-28 days), group B - periovulatory (8-20 days) and group C - postmenopausal. Intra-operative blood loss was calculated during surgery while postoperative blood loss was determined by measuring the amount of drainage. All menstrual data were recorded. Age, body mass index and weight of the resected tissue were also considered as factors that can affect the amount of bleeding and therefore were also evaluated in terms of significance. There was no significant difference between groups regarding intra-operative and postoperative blood loss. Body mass index and weight of the resected tissue had a strong influence on the time of drain removal and the amount of intra-operative and postoperative blood loss. This study strongly suggests that the menstrual period does not affect perioperative blood loss for operations on areas less sensitive to sex hormone levels such as the abdomen. In light of the results, we can conclude that the patient's menstrual date is not of vital importance regarding perioperative blood loss when planning surgery for areas with less sensitivity to menstrual hormonal changes.


Visuoperceptive impairment in adult patients with occipital lobe epilepsies.
The aim of the study was to verify whether adult patients with occipital lobe epilepsy (OLE) are at risk for cognitive impairment compared with controls. Twenty patients with OLE and 20 controls, matched as closely as possible to the epilepsy group in terms of gender, age, education, and intelligence, were administered a battery of tests based on visuoperceptive domains. None of the participants was affected by progressive pathologies, received any medication other than antiepileptic drugs, nor had a psychiatric background. We noted a statistically significant difference between patients with OLE and controls in the following tasks: Perceptive Differences Test, and Object Denomination Test, and Famous Faces Test (P<0.05). No difference was noted between symptomatic and nonsymptomatic patients with respect to neuropsychological results (P>0.05). The linear regression analysis performed did not show any statistically significant contribution by clinical variables. Our data confirm that patients with OLE manifest subtle difficulties in processing and mental manipulation of visual spatial data.


TRANSFORAMINAL INTERSOMATIC LUMBAR ARTHRODESIS: COMPARISON BETWEEN AUTOGRAFT AND CAGE IN PEEK.
To compare the postoperative outcomes of transforaminal intersomatic lumbar arthrodesis with structured iliac bone autograft and PEEK device. The total of 93 medical records of patients undergoing transforaminal intersomatic fusion between January 2012 and July 2017 with at least 1 year of postoperative follow-up, with complete medical record, containing clinical file and radiological exams, were reviewed. From the medical records evaluated, 48 patients underwent the procedure with structured iliac autograft (group 1) and 45 with PEEK device (group 2). There was an improvement in functional capacity in both groups (p < 0.001), however there was no difference when comparing them (p = 0.591). The postoperative clinical and radiological results of lumbar arthrodesis with TLIF technique, using a structured iliac bone autograft compared to a PEEK device, were similar. Level of Evidence II, Retrospective study.


Speciation of heavy metals in biosolids of wastewater treatment plants at Mysore, Karnataka, India.
Urban wastewater treatment leads to the generation of large quantities of biosolids. Accumulation of biosolids is a problem of environmental relevance due to the existence of heavy metals in the biosolids. Determination of total metal in biosolid provides information relating pollution levels. Determination of their mobilization capacity and behaviour in the environment is an important task. An experimental approach commonly used for studying the mobility, transport and bioavailability of metal in biosolids is the use of selective sequential extraction procedure. In the present study an attempt has been made to study the heavy metal properties in biosolid samples collected from urban wastewater treatment plants located at Mysore, Karnataka. Few heavy metals selected for the present study are cadmium, chromium, copper, iron, nickel and zinc. The concentration of these metals in biosolids and their partition in different fractions are studied. The speciation of metals based on the sequential extraction scheme was carried out. The concentration of heavy metals is lower than that established by European legislation. The residual fraction has the maximum percentage of heavy metals whereas, only a small fraction of heavy metals (Fe, Zn and Cd) are extracted in the most soluble fractions, exchangeable and carbonate fractions.


Complete recovery from segmental zoster paresis confirmed by magnetic resonance imaging: A case report.
Segmental zoster paresis, depending on the affected area, can present with severe clinical manifestations and render patients unable to perform activities of daily living. Therefore, it is necessary to diagnose and treat such a condition rapidly. No studies have reported using magnetic resonance imaging (MRI) to identify clinical abnormalities associated with this condition or its complete recovery. This rare case report evaluated the changes in MRI findings before and after the patient's motor symptoms recovered. A 79-year-old woman with a history of rheumatoid arthritis and psoriasis visited the hospital for skin rashes and pain in the C5-T2 segments. The diagnosis was herpes zoster infection, and treatment was initiated. However, motor weakness suddenly occurred 14 d after the initial symptom presentation. We confirmed abnormal findings in the nerves and muscles invaded by the shingles using electromyography and MRI. After 17 mo, the patient's symptoms had completely normalized, and MRI confirmed that there were no abnormalities. MRI can be a useful diagnostic modality for segmental zoster paresis and patient evaluation during recovery from motor complications.


High-efficiency mid-infrared InGaAs/InP arrayed waveguide gratings.
Photonic integrated circuits and mid-infrared quantum cascade lasers have attracted significant attention over the years because of the numerous applications enabled by these compact semiconductor chips. In this paper, we demonstrate low loss passive waveguides and highly efficient arrayed waveguide gratings that can be used, for example, to beam combine infrared (IR) laser arrays. The waveguide structure used consists of an In0.53Ga0.47As core and InP cladding layers. This material system was chosen because of its compatibility with future monolithic integration with quantum cascade lasers. Different photonic circuits were fabricated using standard semiconductor processes, and experiments conducted with these chips demonstrated low-loss waveguides with an estimated propagation loss of ∼ 1.2 dB/cm as well as micro-ring resonators with an intrinsic Q-factor of 174,000. Arrayed waveguide gratings operating in the 5.15-5.34 µm range feature low insertion loss and non-uniformity of ∼ 0.9 dB and ∼ 0.6 dB, respectively. The demonstration of the present photonic circuits paves the path toward monolithic fabrication of compact infrared light sources with advanced functionalities beneficial to many chemical sensing and high-power applications.


Porokeratosis simulating Bowen's disease on dermoscopy.
Porokeratosis is a disorder of epidermal keratinization characterized by the presence of annular hyperkeratotic plaques. Its etiopathogenesis is not yet fully understood, but a relationship with immunosuppression has been reported. Dermoscopic examination revealed a classic yellowish-white ring-like structure that resembled "volcanic crater contour" - the so-called cornoid lamella. We describe a case of porokeratosis in a female patient with chronic lymphedema, which was similar to Bowen's disease due to the many glomerular vessels seen on clinical examination and dermoscopy.


The Deadly Quartet (Covid-19, Old Age, Lung Disease, and Heart Failure) Explains Why Coronavirus-Related Mortality in Northern Italy Was So High.
Since its outbreak in China at the end of 2019, the new coronavirus disease (COVID-19) was characterized by both easy spreading and high mortality. The latter proved to be way more elevated in the North of Italy -with a peak of 18.4% in region Lombardia and even 31% in the city of Bergamo and surrounding county- than in the rest of the world. In an attempt to conceptualize the reasons for such a dramatic situation, four key elements have been identified: COVID-19 itself, old age, lung disease, and heart failure. Their harmful combination has been named "The deadly quartet". The underlying risk factors, among which a lot of them are distinctive features of the population in northern Italy, have been summarized as "unmodifiable", "partially modifiable", and "modifiable", for the sake of clarity. Up-to-date scientific evidence in this field has been described in the form of a narrative and easy-to-read review.


On dose cube pixel spacing pre-processing for features extraction stability in dosiomic studies.
Dosiomics allows to parameterize regions of interest (ROIs) and to produce quantitative dose features encoding the spatial and statistical distribution of radiotherapy dose. The stability of dosiomics features extraction on dose cube pixel spacing variation has been investigated in this study. Based on 17 clinical delivered dose distributions (Pn), dataset has been generated considering all the possible combinations of four dose grid resolutions and two calculation algorithms. Each dose voxel cube has been post-processed considering 4 different dose cube pixel spacing values: 1x1x1, 2x2x2, 3x3x3 mm3 and the one equal to the planning CT. Dosiomics features extraction has been performed from four different ROIs. The stability of each extracted dosiomic feature has been analyzed in terms of coefficient of variation (CV) intraclass correlation coefficient (ICC). The highest CV mean values were observed for PTV ROI and for the grey level size zone matrix features family. On the other hand, the lowest CV mean values have been found for RING ROI for the grey level co-occurrence matrix features family. P3 showed the highest percentage of CV >1 (1.14%) followed by P15 (0.41%), P1 (0.29%) and P13 (0.19%). ICC analysis leads to identify features with an ICC >0.95 that could be considered stable to use in dosiomic studies when different dose cube pixel spacing are considered, especially the features in common among the seventeen plans. Considering the observed variability, dosiomic studies should always provide a report not only on grid resolution and algorithm dose calculation, but also on dose cube pixel spacing.


Furazolidone is a selective in vitro candidate against Leishmania (L.) chagasi: an ultrastructural study.
The current treatment for leishmaniasis is unsatisfactory due to toxic side effects, high cost, and problems with drug resistance. Various approaches have been used to identify novel drug candidates to treat Leishmania sp. parasites including the use of re-purposed drugs. Furazolidone is a nitrofuran derivative with antiprotozoal and antibacterial activity and is used for the treatment of giardiasis. In the present work, we determined the in vitro antileishmanial activity of furazolidone and its ability to induce ultrastructural alterations of parasites. Promastigotes of Leishmania (L.) chagasi, Leishmania (V.) braziliensis, Leishmania (L.) major, and Leishmania (L.) amazonensis were highly susceptible to furazolidone, with IC(50) values ranging between 0.47 and 0.73 microg/mL. Furazolidone was also very effective against L. chagasi intracellular amastigotes, and despite mammalian cytotoxicity, the selectivity index was 8.0 in human monocytes. The drug also had limited toxicity in mice erythrocytes. Furazolidone demonstrated specific activity against Leishmania, a potential consequence of the lack of macrophage nitric oxide activation. As determined by electron transmission microscopy, drug treatment induced severe damage to the parasite mitochondria and nucleus. This older oral drug is an effective agent for the treatment of L. (L.) chagasi in vitro and is a novel candidate for further experimental studies.


RNA synthesised during oogenesis and early embryogenesis in an insect egg (Euscelis plebejus).
RNA labelled during oogenesis or early embryogenesis was isolated from eggs of the leaf hopperEuscelis plebejus. The polyadenylated RNA fraction deposited during early oogenesis accounted for approximately 2.7% of the total RNA content of the newly laid egg. This fraction differed significantly in molecular weight (15-32 S) from poly(A)-containing RNA synthesised between early cleavage and early germ anlage stages (4-20S). Locally injected3H-uridine spread through the egg within approximately 3 h. A considerable fraction (25-35%) of label injected as3H-uridine during early cleavage was recovered in DNA at subsequent stages (10-20 h later); labelled RNA was not found prior to the cellular blastoderm stage. When the yolk-endoplasm was separated from the blastoderm cells, only the latter contained demonstrable amounts of RNA synthesised by the embryo. Of the precursor incorporated into embryonic RNA, approximately 10% was found in the polyadenylated fraction at the early blastoderm stage, but only 3% at the early germ anlage stage. No differences in size distribution of polyadenylated RNA were evident between anterior and posterior halves of the early germ anlage stage.


Direct and indirect organogenesis of Clivia miniata and assessment of DNA methylation changes in various regenerated plantlets.
Clivia miniata is an important indoor ornamental plant and has been reported to have medicinal value. We developed an efficient in vitro micropropagation protocol from young leaves (indirect organogenesis), young petals (indirect organogenesis) and shoot tips (direct organogenesis) of this plant. Using young leaves and shoot tips as explants, the regeneration frequencies were much higher than those in previous investigation and the regeneration was dependent upon less nutrition. We speculated that the leaf-derived callus can generate amino acids necessary for protein synthesis by itself. We employed the methylation-sensitive amplified polymorphism (MSAP) method to assess cytosine methylation variation in various regenerated plantlets and between organs. The MSAP profiles indicated that the frequency of somaclonal variation in the form of cytosine methylation was highest in petal-derived plantlets followed by secondary leaf-derived, primary leaf-derived and shoot tip-derived plantlets, but the methylation variation in petal-derived plantlets was lower than between petals and leaves of a single plant. The results indicated that the methylation variation in regenerated plantlets was related to the types of explants, regeneration pathways and number of regeneration generations. Two possible factors for the highest somaclonal variation rate in petal-derived plantlets are the callus phase and petal-specific set of epigenetic regulators. The property of meristem integrity can account for the lowest variation rate in shoot tip-derived plantlets. Moreover, the secondary plantlets underwent a longer total period of in vitro culture, which can explain why the methylation variation rate in the secondary plantlets is higher than in the primary ones. Methylation variation in regenerated plantlets of C. miniata was found to be related to the types of explants, regeneration pathways and number of regeneration generations.


3d Virtual Histology Reveals Pathological Alterations of Cerebellar Granule Cells in Multiple Sclerosis.
We investigate structural properties of neurons in the granular layer of human cerebellum with respect to their involvement in multiple sclerosis (MS). To this end we analyze data recorded by X-ray phase contrast tomography from tissue samples collected post mortem from a MS and a healthy control group. Using automated segmentation and histogram analysis based on optimal transport theory (OT) we find that the distributions representing nuclear structure in the granular layer move to a more compact nuclear state, i.e. smaller, denser and more heterogeneous nuclei in MS. We have previously made a similar observation for neurons of the dentate gyrus in Alzheimer's disease, suggesting that more compact structure of neuronal nuclei which we attributed to increased levels of heterochromatin, may possibly represent a more general phenomenon of cellular senescence associated with neurodegeneration.


Proto-oncoprotein Zbtb7c and SIRT1 repression: implications in high-fat diet-induced and age-dependent obesity.
Zbtb7c is a proto-oncoprotein that controls the cell cycle and glucose, glutamate, and lipid metabolism. Zbtb7c expression is increased in the liver and white adipose tissues of aging or high-fat diet-fed mice. Knockout or knockdown of Zbtb7c gene expression inhibits the adipocyte differentiation of 3T3-L1 cells and decreases adipose tissue mass in aging mice. We found that Zbtb7c was a potent transcriptional repressor of SIRT1 and that SIRT1 was derepressed in various tissues of Zbtb7c-KO mice. Mechanistically, Zbtb7c interacted with p53 and bound to the proximal promoter p53RE1 and p53RE2 to repress the SIRT1 gene, in which p53RE2 was particularly critical. Zbtb7c induced p53 to interact with the corepressor mSin3A-HADC1 complex at p53RE. By repressing the SIRT1 gene, Zbtb7c increased the acetylation of Pgc-1α and Pparγ, which resulted in repression or activation of Pgc-1α or Pparγ target genes involved in lipid metabolism. Our study provides a molecular target that can overexpress SIRT1 protein in the liver, pancreas, and adipose tissues, which can be beneficial in the treatment of diabetes, obesity, longevity, etc.


An automated high pressure reaction vessel for routine preparation of short-lived radiopharmaceuticals.
A remote, automated high-pressure/temperature system for preparation of radiopharmaceuticals is described. The system was routinely employed in our laboratories in the production of multimillicurie amounts of 11C labeled D,L-amino acids in more than 90 production runs, with typical final D,L-product activities of 240-400 mCi. The high pressure system, however, is not restricted only to the production of 11C labeled amino acids, but for application in any synthetic procedure requiring high pressure (approximately 300 psia) and temperature (greater than 300 degrees C).


Effect of dexfenfluramine on metabolic and neurochemical measures in restraint-stressed ob/ob mice.
Genetically obese (ob/ob) and lean C57BL/6J mice were subjected to 1 h acute restraint stress, and the effects on plasma glucose and corticosterone, and brain amines, were assessed relative to nonrestrained groups. Some mice were treated with dexfenfluramine (DFEN) prior to restraint. Compared with their lean counterparts, ob/ob mice had elevated basal plasma levels of both glucose and corticosterone. Both lean and ob/ob showed comparable absolute restraint-induced increments in these measures, but the fractional increase was much smaller in ob/ob. Metabolism of NE, DA and 5-HT was also assessed from the ratio of the metabolite-to-amine in various brain regions. Obese mice had smaller restraint-related increases in monoamine metabolism than lean their counterparts. Acute DFEN increased corticosterone, but tended to reduce the restraint-induced elevations in glucose and monoamine metabolism.


Reconstruction of mitochondrial genomes from raw sequencing data provides insights on the phylogeny of Ixodes ticks and cautions for species misidentification.
High-throughput sequencing (HTS) technology has profoundly been involved in sequencing whole genomes of several organisms in a fast and cost-effective manner. Although HTS provides an alternative biomonitoring method to the time-consuming and taxonomy-expertise dependent morphological approach, still we cannot rule out the possibility of the impediment and misidentification biases. In this article we aim to retrieve whole mitochondrial genome (mitogenome) sequences from publicly available raw sequencing data for phylogenetic comparison of Ixodes persulcatus. For this comparison, we sequenced whole mitogenomes of four I. persulcatus ticks from Japan and constructed mitogenomes from raw sequencing data of 74 I. persulcatus ticks from China. Bayesian phylogenetic trees were inferred by the concatenated fifteen mitochondrial genes. We further tested our results by the phylogenetic analysis of cytochrome c oxidase subunit 1 (cox1) gene and internal transcribed spacer 2 (ITS2) sequences. Our findings showed that 70 constructed mitogenomes from China were clustered with the sequenced four mitogenomes of I. persulcatus from Japan. We also revealed that mitogenome sequences retrieved from two data sets CRR142297 and CRR142298 were clustered with Ixodes nipponensis. Moreover, other two mitogenome sequences from CRR142310 and CRR142311 formed a clade with Ixodes pavlovskyi. The phylogenetic analysis of cox1 gene and ITS2 sequences confirmed the identification errors of these four samples. The overall phylogenetics in our study concluded that accurate morphological identification is necessary before implementing HTS to avoid any misidentification biases.


A clinic-based motivational intervention improves condom use among subgroups of youth living with HIV.
More than 50% of youth living with HIV (YLH) have unprotected sex. In previous studies, we reported effects of a motivational interviewing-based multirisk reduction intervention, "Healthy Choices" in improving motivation, depression, and viral load in YLH. In this study, we report the effect of the intervention on increasing condom use. Six waves of longitudinal data (n = 142) across a period from baseline through 15 months postintervention were analyzed. The developmental trajectory modeling method was used for program effect evaluation. The three groups detected with distinct sexual risks were: Persistent low sexual risk (PLSR), delayed high sexual risk, and high and growing sexual risk with regard to levels and time trajectories of condom use throughout the trial. Receiving Healthy Choices increased the likelihood to be in the PLSR group (63% vs. 32%, p < .01) and reduced the likelihood to be in the delayed high sexual risk group (16% vs. 50%, p < .05). Receiving the intervention was also associated with progressive reductions in no-condom sex for PLSR youth (adjusted β = -.325, p < .01) and high and growing sexual risk youth (adjusted β = -.364, p < .01). The motivational interviewing-based program Healthy Choices, when delivered in clinic settings, can prevent unprotected sex in subgroups of YLH, although more intensive interventions may be needed to change risk trajectories among those at highest risk of transmitting the AIDS virus. Developmental trajectory analysis provides an alternative approach to evaluate program effects for study samples that contain distinct subgroups.


ASA classification and surgical severity grading used to identify a high-risk population, a multicenter prospective cohort study in Swedish tertiary hospitals.
Identification of surgical populations at high risk for negative outcomes is needed for clinical and research purposes. We hypothesized that combining two classification systems, ASA (American Society of Anesthesiology physical status) and surgical severity, we could identify a high-risk population before surgery. We aimed to describe postoperative outcomes in a population selected by these two classifications system. Data were collected in a Swedish multicentre, time-interrupted prospective, consecutive cohort study. Eligibility criteria were age ≥18 years, ASA ≥3, elective or emergent, major to Xmajor/complex (Specialist Procedure Codes used in United Kingdom), gastrointestinal, urogenital or orthopaedic procedures. Postoperative morbidity was identified by the Postoperative Morbidity Survey on postoperative days 3 ± 1, 7 ± 1, 10 + 5 and graded for severity by the Clavien-Dindo system. Mortality was assessed at 30, 180 and 360 days. Postoperative morbidity was 78/48/47 per cent on postoperative days 3/7/10. Majority of morbidities (67.5 per cent) were graded as >1 by Clavien-Dindo. Any type of postoperative morbidity graded >1 was associated with increased risk for death up to one year. The mortality was 5.7 per cent (61/1063) at 30 days, 13.3 per cent (142/1063) at 6 months and 19.1 per cent (160/1063) at 12 months. Severity classification as major to Xmajor/complex and ASA ≥3 could be used to identify a high-risk surgical population concerning postoperative morbidity and mortality before surgery. Combining the two systems future electronic data extraction is possible of a high-risk population in tertiary hospitals.


Comparative cytotoxicity of carmustine (BCNU), nimustine (ACNU) and elmustine (HeCNU) after depletion of O6-alkylguanine-DNA alkyltransferase (O6-AGT).
BCNU was reported to have about a 6- to 8- fold lower cytotoxic potency than ACNU in cell lines naturally deficient in O6-AGT. In seven tumor cell lines with an O6-AGT activity ranging from 40 to 360 fmol/mg the cytotoxic potency of BCNU, ACNU and HeCNU, without and after O6-AGT depletion by O6-BG, was determined. Without O6-AGT depletion, BCNU was superior to both other drugs in tumor cells with high O6-AGT activity. After O6-AGT depletion, the cytotoxic potency (comparison of IC50 values) of ACNU was higher than that of BCNU (p=0.016) or that of HeCNU (p=0.016) in all tumor cell lines. We conclude that (without O6-AGT depletion) BCNU is the drug of choice especially in tumor cells with high transferase activity. The higher cytotoxic potency of ACNU after O6-AGT depletion as compared to BCNU after O6-AGT depletion is countered by the higher toxicity of ACNU in patients necessitating a clinical dose reduction as compared to BCNU. Thus, we would not expect superiority of ACNU + O6-BG over BCNU+ O6-BG after systemic administration.


Hexose transport in microvascular endothelial cells cultured from bovine retina.
The nature of glucose transport at the microvascular blood-retinal barrier was studied using primary cultures of microvascular endothelial cells from bovine retina. Uptake of 3-O-methyl-D-glucose (3MG), a non-metabolizable glucose analogue, was rapid and equilibrative. 3MG uptake could be inhibited by traditional glucose transport inhibitors such as phloretin, phlorizin and cytochalasin B but not by agents that deplete intracellular ATP (2,4-dinitrophenol) or that abolish the sodium gradient (ouabain). Uptake of 3MG by the cells could be stimulated by preloading with 50 mM-glucose, a phenomenon known as counter-transport. Insulin had no effect on 3MG uptake in the cells even after prolonged insulin deprivation. These results demonstrate the existence of a facilitative-diffusion type of glucose transport system in endothelial cells of the retinal microvasculature. Studies with 2-deoxy-D-glucose demonstrated that transport was not rate limiting for metabolism and that the endothelial cells contain free sugar when exposed to an extracellular sugar concentration in the physiologic range. The presence of this free sugar within the cell is necessary for efficient transendothelial transfer of glucose from blood to retina but it could also provide the basis for capillary damage in diabetes mellitus.


Cell lines from the soft tick Ornithodoros moubata.
Primary cell cultures (n = 16) were initiated from tissues of embryonic and neonatal larval Ornithodoros moubata following methods developed for hard ticks. After maintenance for 20-25 months in vitro, cell multiplication commenced in surviving cultures, leading to the establishment of six cell lines designated OME/CTVM21, 22, 24, 25, 26 and 27. All lines are maintained at 28 degrees C, with subculture at 2-8 week intervals. The cultures comprise heterogeneous populations of large cells of 15-100 microm in diameter, often with finger-like protrusions and/or intracellular crystals, rarely attached, predominantly floating and forming clumps or hollow multicellular vesicles up to 1 mm in diameter. Attempts to cryopreserve the cells are described. Tick-borne encephalitis virus has been serially passaged ten times in OME/CTVM21 cells without significant decrease in virus production and with no change in its biological properties as shown by the size and morphology of plaques produced in porcine kidney cells.


Water level of inland saline wetlands with implications for CO2 and CH4 fluxes during the autumn freeze-thaw period in Northeast China.
Zhalong wetland is the largest inland saline wetland in Asia and susceptible to imbalance and frequent flooding during the freeze-thaw period. Changes in water level and temperature can alter the rate of greenhouse gas release from wetlands and have the potential to alter Earth's carbon budget. However, there are few reports on how water level, temperature, and their interactions affect greenhouse gas flux in inland saline wetland during the freeze-thaw period. This study revealed the characteristics of CO2 and CH4 fluxes in Zhalong saline wetlands at different water levels during the autumn freeze-thaw period and clarifies the response of CO2 and CH4 fluxes to water levels. The significance analysis of cumulative CO2 fluxes at different water levels showed that water levels did not have a significant effect on cumulative CO2 release fluxes from wetlands. Water levels, temperature, soil moisture content, soil nitrate, and ammonium nitrogen content and organic carbon content could explain 24.5-98.9% of CO2 and CH4 flux variation. There were significant differences in the average and cumulative CH4 fluxes at different water levels. The higher the water levels, the higher the CH4 fluxes. In short, water level had a significant effect on wetland methane fluxes, but not on carbon dioxide fluxes.


Lower urinary tract symptoms in adults treated for posterior urethral valves in childhood: matched cohort study.
We compared the prevalence and bother of lower urinary tract symptoms in adults treated for posterior urethral valves in childhood and population based controls. Questionnaires were mailed to patients 18 years and older treated at our institution for posterior urethral valves. Of the 124 patients 68 (64.2%) participated, 18 were unavailable and 38 did not participate. Age and sex matched controls were randomly identified from a population based study (response proportion 62.4%). Danish Prostatic Symptom Score was used to assess occurrence (never/rarely/often/always) and bother (none/small/moderate/major) of 12 different lower urinary tract symptoms. Chi-square test was used for the analyses stratified by patient/control status, with the 4-point ordinal scale as the outcome. Median age of 68 patients with posterior urethral valves and 272 controls (ratio 1:4) was 38.5 years (range 18 to 57). Overall, at least 1 moderate or severe lower urinary tract symptom was reported by 32.4% of patients with posterior urethral valves and 15.8% of controls (p=0.002). Mild hesitancy, weak stream, incomplete emptying and straining were reported twice as often by patients with posterior urethral valves as by controls (p<0.05). Prevalence of any urgency incontinence (14.7% vs 4.8%, p=0.014) and any stress incontinence (11.8% vs 3.0%, p=0.005) was increased at least 3-fold in patients with posterior urethral valves compared to controls. Furthermore, prevalence of bother from several lower urinary tract symptoms was increased approximately 2-fold in patients with posterior urethral valves compared to controls (p<0.05). However, most patients and controls reported no or small bother. In adulthood the occurrence and bother of most lower urinary tract symptoms are increased approximately 2-fold in patients with posterior urethral valves compared to the general population. However, in this cohort of young and middle-aged men most symptoms are mild.


Implementing Universal Suicide Risk Screening in a Pediatric Hospital.
Health care providers are in a prime position to identify teens at risk for suicide, yet many do not. The research team developed and implemented a hospitalwide program to identify teens at elevated risk for suicide and connect them with services. Screening was implemented at both locations of a pediatric hospital, including two emergency departments, three urgent care clinics, and ambulatory clinics. Patients aged 12 years and older presenting for care were screened for suicide risk using the Ask Suicide-Screening Questions (ASQ) in most settings, while the Columbia-Suicide Severity Rating Scale (C-SSRS) was used in mental health areas. A social worker responded to positive screens to complete a more thorough assessment and determine next steps. Social workers also completed outreach to patients in the weeks following a positive screen. Implementation began with pilot locations and expanded after refinements were made. Stakeholders provided screening recommendations, and education was provided prior to implementation. The cost of implementation was calculated based on the time screening required from nursing and social work. Review of the program focused on implementation fidelity, quality improvement, and trends among screening results. During the first year of screening, 138,598 screens were completed, and 6.8% of screens were positive for elevated risk. The annualized cost of the program was estimated to be $887,708.65 for personnel directly involved in screening and following up on positive screens. Early involvement of stakeholders and hospital leaders and a robust response plan were essential to successful implementation of this suicide-screening program.


Three-dimensional reconstruction of median eminence microvascular modules.
To test the hypothesis that the median eminence microvasculature has a direct regulatory role in the hormonal communication between the brain and the pituitary gland, it is necessary to determine whether the physical means for such control (e.g. smooth muscle sphincters strategically located in the capillary plexus) actually exists. Our approach is to search for such structures in transmission electron micrographs of thin serial sections of the median eminence. The complexity of these images and the anticipated need to include large numbers of them in the study led us to consider computer reconstruction for this problem. We report here the successful three-dimensional reconstruction of capillary modules using digital image processing techniques for capillary feature detection/extraction, for construction of montages (mosaics) of overlapping images of the same section, and for automatic image registration by two independent methods without the use of fiducial marks. These tasks have been performed manually in nearly all the published neurobiological reconstructions; here they are performed by programs using only the mathematical properties of the images. Methods like those described here provide the only practical means for executing large scale reconstructions and gaining significant new information about the regulation of blood flow in this region of the brain.


Effects of Impurity Doping on the Luminescence Performance of Mn4+-Doped Aluminates with the Magnetoplumbite-Type Structure for Plant Cultivation.
Mn4+ activated LaMgAl11O19 (LMA/Mn4+) with red emitting phosphor was obtained by sintering under air conditioning. The X-ray diffraction pattern Rietveld refinement results reveal that three six-fold coordinated Al sites are substituted by Mn4+ ions. Furthermore, the chemical valence state of manganese in the LMA host was further confirmed through X-ray photoelectron spectroscopy (XPS) and electron paramagnetic resonance (EPR). Photoluminescence emission (PL) and excitation (PLE) spectra of LMA/Mn4+ as well as the lifetime were measured, and the 663 nm emission is ascribed to the ²Eg→⁴A2g from the 3d³ electrons in the [MnO₆]8- octahedral complex. The emission spectrum matches well with the absorption of phytochrome. Temperature-dependent PL spectra show that the color changes of the phosphor at 420 K are 0.0110 for Δx and -0.0109 for Δy. Moreover, doping Zn2+ and Mg2+ ions in the host enhances the emission intensity of Mn4+ ions. These results highlight the potential of LMA/Mn4+ phosphor for a light-emitting diode (LED) plant lamp.


Altered self-assembly and apatite binding of amelogenin induced by N-terminal proline mutation.
A single Pro-70 to Thr (p.P70T) mutation of amelogenin is known to result in hypomineralised amelogenesis imperfecta (AI). This study aims to test the hypothesis that the given mutation affects the self-assembly of amelogenin molecules and impairs their ability to conduct the growth of apatite crystals. Recombinant human full-length wild-type (rh174) and p.P70T mutated amelogenins were analysed using dynamic light scattering (DLS), protein quantification assay and atomic force microscopy (AFM) before and after the binding of amelogenins to hydroxyapatite crystals. The crystal growth modulated by both amelogenins in a dynamic titration system was observed using AFM. As compared to rh174 amelogenin, p.P70T mutant displayed significantly increased sizes of the assemblies, higher binding affinity to apatite, and decreased crystal height. Pro-70 plays an important structural role in the biologically relevant amelogenin self-assembly. The disturbed regularity of amelogenin nanospheres by this single mutation resulted in an increased binding to apatite and inhibited crystal growth.


[Effect of ginsenoside Rb1 on insulin signal transduction pathway in hippocampal neurons of high-glucose-fed rats].
To study the effect of ginsenoside Rb1 on GSKbeta/IDE signal transduction pathway and Abeta protein secretion in hippocampal neurons of high glucose-treated rats. Hippocampal neurons of 24 h-old newly born SD rats were primarily cultured, inoculated in culture medium under different conditions, and then divided into the normal group, the high glucose group, the LiCl group and the Rb1 group. After being cultured for 72 h, the expressions of their phosphorylated GSK3beta, total GSK3beta and IDE protein were detected by Western blotting analysis. The mRNA expressions of GSK3beta and IDE were determined by RT-PCR. The ELISA assay was used to detect the secretion of Abeta protein in cell supernatant. Compared with the normal group, the high glucose group showed increase in the p/tGSK3beta protein ratio and the secretion of Abeta protein and decrease in IDE protein and mRNA (P < 0.05). Compared with the high glucose group, both Rb1 and LiCl groups showed decrease in the p/tGSK3beta protein ratio and the expression of Abeta protein and increase in IDE protein and mRNA expression (P < 0.05). Compared with the LiCl group, the Rb1 group showed no significant difference in the expressions of p/tGSK3beta protein, IDE protein, mRNA and Abeta protein expression. In addition, the GSK3beta mRNA expression of the four groups had no significant difference. Ginsenoside Rb1 may reduce the secretion of Abeta protein in hippocampal neurons by reducing the phosphorylation of GSK3beta, down-regulating the ratio of pGSK3beta/GSK3beta and upregulating the expression of IDE.


Prognostic value of miR-219-5p in relation to mortality in patients with small cell lung cancer: a retrospective, observational cohort study in China.
Small cell lung cancer (SCLC) is a lethal human malignancy, and previous studies support the contribution of microRNA to cancer progression. The prognostic value of miR-219-5p in patients with SCLC remains unclear. This study aimed to evaluate the predictive value of miR-219-5p with respect to mortality in patients with SCLC and to incorporate miR-219-5p level into a prediction model and nomogram for mortality. Retrospective observational cohort study. Our main cohort included data from 133 patients with SCLC between 1 March 2010 and 1 June 2015 from the Suzhou Xiangcheng People's Hospital. Data from 86 patients with non-SCLC at Sichuan Cancer Hospital and the First Affiliated Hospital of Soochow University were used for external validation. Tissue samples were taken during admission and stored, and miR-219-5p levels were measured at a later date. A Cox proportional hazard model was used for survival analyses and for analysing risk factors to create a nomogram for mortality prediction. The accuracy of the model was evaluated by C-index and calibration curve. Mortality in patients with a high level of miR-219-5p (≥1.50) (n=67) was 74.6%, while mortality in the low-level group (n=66) was 100.0%. Based on univariate analysis, we included significant factors (p<0.05) in a multivariate regression model: patients with high level of miR-219-5p (HR 0.39, 95% CI 0.26-0.59, p<0.001), immunotherapy (HR 0.44, 95% CI 0.23-0.84, p<0.001) and prognostic nutritional index score >47.9 (HR=0.45, 95% CI 0.24-0.83, p=0.01) remained statistically significant factors for improved overall survival. The nomogram had good accuracy in estimating the risk, with a bootstrap-corrected C-index of 0.691. External validation indicated an area under the curve of 0.749 (0.709-0.788). The miR-219-5p level was associated with a reduced risk of mortality in patients with SCLC. A nomogram incorporating MiR-219-5p level and clinical factors demonstrated good accuracy in estimating the risk of overall mortality. Prospective validation of the prognostic nomogram is needed.


[The dynamics of thyroid diseases in the inhabitants of Kiev and Kiev Province after the accident at the Chernobyl Atomic Electric Power Station].
Autoimmune affections of the thyroid gland were found to be on the increase as were Hashimoto's disease and follicular adenoma, as evidenced by a pathomorphic study in surgically removed thyroid glands of 12684 patients over 1984-1993. Rise in the frequency of malignant tumors was moderate. Increase in the incidence of papillary microcarcinoma against the background of another surgical pathology of the thyroid gland may be an unfavourable prognostic sign of further growth of morbidity of clinically manifest forms of carcinoma in Kiev and the Kiev Province.


Sarcoid involvement of the supraorbital nerve: MR and histologic findings.
We report the MR and histologic findings of neurosarcoidosis presenting as a mass involving the supraorbital nerve in a 29-year-old woman in whom this was the first manifestation of the disease. The features and associations of neurosarcoidosis and the response to treatment are discussed.


Diabetic retinopathy screening and treatment through the Brazilian National Health Insurance.
The current study aimed to investigate diabetic retinopathy (DR) screening and treatment coverages among diabetic patients evaluated through the Brazilian National Health Insurance from 2014 to 2019. The Brazilian Public Health System Information Database was used as the primary data source. DR screening coverage was calculated as the rate of procedures of clinical dilated fundus exam and color fundus photograph over the number of diabetic patients. DR treatment coverage was calculated as the rate of procedures of intravitreal injection, photocoagulation, and panretinal photocoagulation over the number of diabetic patients presumably in need of DR treatment. The overall screening coverage increased from 12.1% in 2014 to 21.2% in 2019 (p < 0.001) with substantial regional discrepancies so that North region was the only one with no changes along the period. The overall treatment coverage increased from 27.7% in 2014 to 44.1% in 2019, with Southeast and Midwest absorbing the demand for service from the North, Northeast and South. Despite an improvement along the past years, both screening and treatment coverages for DR in diabetes patients are ineffective in Brazil. Public health policies should address resources disparities throughout the country aiming to offer same healthcare conditions to patients regardless their geographic location.


Seizure Reduction with Fluoxetine in Dravet Syndrome.
An adult woman with Dravet syndrome (documented SCN1A mutation) experienced a marked reduction in seizures when treated with the selective serotonin reuptake inhibitor (SSRI) fluoxetine. The seizure reduction may be partly to reductions associated with aging in Dravet patients, but it appears to be due at least in part to the fluoxetine. A prior preliminary study reported that fenfluramine reduces seizures in patients with Dravet syndrome. Fenfluramine may produce this effect by increasing serotonin brain levels, and SSRIs have been found to possess antiepileptic properties in animal models of epilepsy. Given the known cardiac risks of fenfluramine, consideration of randomized clinical trials with SSRIs should be considered in Dravet syndrome and other epilepsies.


Human papilloma virus E7 oncoprotein abrogates the p53-p21-DREAM pathway.
High risk human papilloma viruses cause several types of cancer. The HPV oncoproteins E6 and E7 are essential for oncogenic cell transformation. E6 mediates the degradation of the tumor suppressor p53, and E7 can form complexes with the retinoblastoma pRB tumor suppressor. Recently, it has been shown that HPV E7 can also interfere with the function of the DREAM transcriptional repressor complex. Disruption of DREAM-dependent transcriptional repression leads to untimely early expression of central cell cycle regulators. The p53-p21-DREAM pathway represents one important means of cell cycle checkpoint activation by p53. By activating this pathway, p53 can downregulate transcription of genes controlled by DREAM. Here, we present a genome-wide ranked list of genes deregulated by HPV E7 expression and relate it to datasets of cell cycle genes and DREAM targets. We find that DREAM targets are generally deregulated after E7 expression. Furthermore, our analysis shows that p53-dependent downregulation of DREAM targets is abrogated when HPV E7 is expressed. Thus, p53 checkpoint control is impaired by HPV E7 independently of E6. In summary, our analysis reveals that disruption of DREAM through the HPV E7 oncoprotein upregulates most, if not all, cell cycle genes and impairs p53's control of cell cycle checkpoints.


Breastfeeding, first-food systems and corporate power: a case study on the market and political practices of the transnational baby food industry and public health resistance in the Philippines.
The aggressive marketing of breastmilk substitutes (BMS) reduces breastfeeding, and harms child and maternal health globally. Yet forty years after the World Health Assembly adopted the International Code of Marketing of Breast-milk Substitutes (The Code), many countries are still to fully implement its provisions into national law. Furthermore, despite The Code, commercial milk formula (CMF) markets have markedly expanded. In this paper, we adopt the Philippines as a case study to understand the battle for national Code implementation. In particular, we investigate the market and political strategies used by the baby food industry to shape the country's 'first-food system', and in doing so, promote and sustain CMF consumption. We further investigate how breastfeeding coalitions and advocates have resisted these strategies, and generated political commitment for a world-leading breastfeeding policy framework and protection law (the 'Milk Code'). We used a case study design and process tracing method, drawing from documentary and interview data. The decline in breastfeeding in the Philippines in the mid-twentieth Century associated with intensive BMS marketing via health systems and consumer advertising. As regulations tightened, the industry more aggressively promoted CMFs for older infants and young children, thereby 'marketing around' the Milk Code. It established front groups to implement political strategies intended to weaken the country's breastfeeding policy framework while also fostering a favourable image. This included lobbying government officials and international organizations, emphasising its economic importance and threats to foreign investment and trade, direct litigation against the government, messaging that framed marketing in terms of women's choice and empowerment, and forging partnerships. A resurgence in breastfeeding from the mid-1980s onwards reflected strengthening political commitment for a national breastfeeding policy framework and Milk Code, resulting in-turn, from collective actions by breastfeeding coalitions, advocates and mothers. The Philippines illustrates the continuing battle for worldwide Code implementation, and in particular, how the baby food industry uses and adapts its market and political practices to promote and sustain CMF markets. Our results demonstrate that this industry's political practices require much greater scrutiny. Furthermore, that mobilizing breastfeeding coalitions, advocacy groups and mothers is crucial to continually strengthen and protect national breastfeeding policy frameworks and Code implementation.


Injection treatment for vascular flares.
Small or minor varicosities pose a considerable cosmetic problem to women afflicted with them. Previous methods of treatment have been either painful, or ineffective, or both. A simplified method of injection of these varices is described.


Non-random distribution of instability-associated chromosomal rearrangement breakpoints in human lymphoblastoid cells.
Genomic instability is observed in tumors and in a large fraction of the progeny surviving irradiation. One of the best-characterized phenotypic manifestations of genomic instability is delayed chromosome aberrations. Our working hypothesis for the current study was that if genomic instability is in part attributable to cis mechanisms, we should observe a non-random distribution of chromosomes or sites involved in instability-associated rearrangements, regardless of radiation quality, dose, or trans factor expression. We report here the karyotypic examination of 296 instability-associated chromosomal rearrangement breaksites (IACRB) from 118 unstable TK6 human B lymphoblast, and isogenic derivative, clones. When we tested whether IACRB were distributed across the chromosomes based on target size, a significant non-random distribution was evident (p<0.00001), and three IACRB hotspots (chromosomes 11, 12, and 22) and one IACRB coldspot (chromosome 2) were identified. Statistical analysis at the chromosomal band-level identified four IACRB hotspots accounting for 20% of all instability-associated breaks, two of which account for over 14% of all IACRB. Further, analysis of independent clones provided evidence within 14 individual clones of IACRB clustering at the chromosomal band level, suggesting a predisposition for further breaks after an initial break at some chromosomal bands. All of these events, independently, or when taken together, were highly unlikely to have occurred by chance (p<0.000001). These IACRB band-level cluster hotspots were observed independent of radiation quality, dose, or cellular p53 status. The non-random distribution of instability-associated chromosomal rearrangements described here significantly differs from the distribution that was observed in a first-division post-irradiation metaphase analysis (p=0.0004). Taken together, these results suggest that genomic instability may be in part driven by chromosomal cis mechanisms.


Detection of adverse events in general surgery using the " Trigger Tool" methodology.
Surgery is one of the high-risk areas for the occurrence of adverse events (AE). The purpose of this study is to know the percentage of hospitalisation-related AE that are detected by the «Global Trigger Tool» methodology in surgical patients, their characteristics and the tool validity. Retrospective, observational study on patients admitted to a general surgery department, who underwent a surgical operation in a third level hospital during the year 2012. The identification of AE was carried out by patient record review using an adaptation of «Global Trigger Tool» methodology. Once an AE was identified, a harm category was assigned, including the grade in which the AE could have been avoided and its relation with the surgical procedure. The prevalence of AE was 36,8%. There were 0,5 AE per patient. 56,2% were deemed preventable. 69,3% were directly related to the surgical procedure. The tool had a sensitivity of 86% and a specificity of 93,6%. The positive predictive value was 89% and the negative predictive value 92%. Prevalence of AE is greater than the estimate of other studies. In most cases the AE detected were related to the surgical procedure and more than half were also preventable. The adapted «Global Trigger Tool» methodology has demonstrated to be highly effective and efficient for detecting AE in surgical patients, identifying all the serious AE with few false negative results.


Exploring needs, barriers to, and facilitators of rehabilitation exercise following revision hip replacement - A grounded theory study.
Evidence on rehabilitation after revision total hip replacement (THR) is inadequate and development of rehabilitation interventions is warranted. Even so, little is known about patients' experiences with revision THR rehabilitation. This study aimed to explore patients' rehabilitation exercise experiences after revision THR. Using constructivist grounded theory, we conducted semi-structured qualitative interviews with twelve patients with completed or almost completed rehabilitation exercise after revision THR. Data collection and analysis were a constant comparative process conducted in three phases; initial, focused, and theoretical. From the data, we generated a substantial theory of the participant's circumstances and ability to integrate rehabilitation exercise into their everyday life after revision THR. Four categories were constructed based on patients' experiences in different contexts: hesitance, fear avoidance, self-commitment, and fidelity. This study highlighted that patients' expectations, past experiences, attitudes, trusts, and circumstances interact to influence engagement and adherence to rehabilitation exercise and described four categories relating to the integration of revision THR rehabilitation exercise into their everyday life. Clinicians should be aware of and account for these categories during rehabilitation exercise. Tailored individual rehabilitation exercise interventions and clinician approaches to optimize engagement and adherence are needed among patients with revision THR.


Positive geographic correlation between soldiers' weapon size and defensive prowess in a eusocial aphid, Ceratovacuna japonica.
Some aphid species produce a soldier caste with enlarged forelegs and horns (weapons). It has been hypothesised that the evolution of morphological specialization by soldiers in social aphids is accelerated by high predation pressure, but this possibility has not been tested. Here, we investigated the relationship between local predator abundance and soldiers' weapon size and aggressiveness in a prey-predator system comprising a eusocial aphid, Ceratovacuna japonica, and its predators (larvae of the butterfly Taraka hamada and of the moth Atkinsonia ignipicta) in two populations with different predator abundances. We found that the soldiers in the predator-abundant population had larger weapons and were more aggressive than those in the population with lower predator abundance. Furthermore, the soldiers' defensive prowess (evaluated as the survival of aphids in the presence of predators) was greater in the predator-abundant population. These results provide the first evidence that a population of eusocial aphids experiencing high predation pressure has soldiers with pronounced defensive traits and defensive prowess.


COVID-19 pandemic and social distancing: economic, psychological, family, and technological effects.
The concept of social isolation is currently understood as a measure of epidemiological containment that aims to reduce the speed of spread of the disease, enabling health services to prepare their resources to cope with the likely increase in demand, while also seeking to provide additional protection to groups considered to be at higher risk. The present narrative review aims to compile and synthesize the literature related to social isolation produced during the COVID-19 pandemic in 2020. This study is a narrative review of the literature on social isolation in the context of the COVID- 19 pandemic. 73 publications were included for full-text reading and were classified into the following categories: levels of social isolation, economic effects, family relationships, health system, mental health of the population, and use of technology. It is necessary to plan an escalation of responses to the consequences of the pandemic, especially in view of the increased demand on the health sector and social services. The negative effects of social isolation can be prevented by public policies that offer a response to the economic recession, maintenance of social work, encouragement of quality care in mental health services, and community support for vulnerable families.


Synthesis of 5-thiodidehydropyranylcytosine derivatives as potential anti-HIV agents.
As a part of our ongoing efforts to identify new anti-HIV agents, a 5'-thiopyrano-nucleoside derivative 4, designed based on 4'-thioD4C 1 and cyclohexenylnucleoside 3, was synthesized. The dihydrothiopyran skeleton of 4 was constructed by the ring closing metathesis of 21 which was synthesized from but-2-yne-1,4-diol. After converting the protecting group from MOM to TBS followed by oxidation, a Pummerer-type thioglycosylation reaction of 24 with persilylated uracil gave the desired 5-thiodihydrothiopyranyluracils 25 and 26 as a mixture of anomers. The conversion of 25 to a cytosine derivative and subsequent deprotection gave a 5-thiodidehydropyranosylcytosine derivative 4 in good yield. The anti-HIV activity of 4 was also evaluated.


Brpf1, a subunit of the MOZ histone acetyl transferase complex, maintains expression of anterior and posterior Hox genes for proper patterning of craniofacial and caudal skeletons.
The epigenetic mechanism involving chromatin modification plays a critical role in the maintenance of the expression of Hox genes. Here, we characterize a mutant of the medaka fish, named biaxial symmetries (bis), in which brpf1, a subunit of the MOZ histone acetyl transferase (HAT) complex, is mutated. The bis mutant displayed patterning defects both in the anterior-posterior axis of the craniofacial skeleton and the dorsal-ventral axis of the caudal one. In the anterior region, the bis mutant exhibited craniofacial cartilage homeosis. The expression of Hox genes was decreased in the pharyngeal arches, suggesting that the pharyngeal segmental identities were altered in the bis mutant. In the posterior region, the bis mutant exhibited abnormal patterning of the caudal skeleton, which ectopically formed at the dorsal side of the caudal fin. The expression of Zic genes was decreased at the posterior region, suggesting that the dorsal-ventral axis formation of the posterior trunk was disrupted in the bis mutant. We also found that the MOZ-deficient mice exhibited an abnormal patterning of their craniofacial and cervical skeletons and a decrease of Hox transcripts. We propose a common role of the MOZ HAT complex in vertebrates, a complex which is required for the proper patterning for skeletal development.


Tumor and Peripheral Immune Status in Soft Tissue Sarcoma: Implications for Immunotherapy.
Soft Tissue Sarcomas (STS) are a heterogeneous and rare group of tumors. Immune cells, soluble factors, and immune checkpoints are key elements of the complex tumor microenvironment. Monitoring these elements could be used to predict the outcome of the disease, the response to therapy, and lead to the development of new immunotherapeutic approaches. Tumor-infiltrating B cells, Natural Killer (NK) cells, tumor-associated neutrophils (TANs), and dendritic cells (DCs) were associated with a better outcome. On the contrary, tumor-associated macrophages (TAMs) were correlated with a poor outcome. The evaluation of peripheral blood immunological status in STS could also be important and is still underexplored. The increased lymphocyte-to-monocyte ratio (LMR) and neutrophil-to-lymphocyte ratio (NLR), higher levels of monocytic myeloid-derived suppressor cells (M-MDSCs), and Tim-3 positive CD8 T cells appear to be negative prognostic markers. Meanwhile, NKG2D-positive CD8 T cells were correlated with a better outcome. Some soluble factors, such as cytokines, chemokines, growth factors, and immune checkpoints were associated with the prognosis. Similarly, the expression of immune-related genes in STS was also reviewed. Despite these efforts, only very little is known, and much research is still needed to clarify the role of the immune system in STS.


Cell responses and hemocompatibility of g-HA/PLA composites.
The objective of this study was to investigate the hemocompatibility and cell responses to some novel poly(L-lactide) (PLA) composites containing surface modified hydroxyapatite particles for potential applications as a bone substitute material. The surface of hydroxyapatite (HA) particles was first grafted with L-lactic acid oligomers to form grafted HA (g-HA) particles. The g-HA particles were further blended with PLA to prepare g-HA/PLA composites. Our previous study has shown significant improvement in tensile properties of these materials due to the enhanced interfacial adhesion between the polymer matrix and HA particles. To further investigate the potential applications of these composites in bone repair and other orthopedic surgeries, a series of in vitro and in vivo experiments were conducted to examine the cell responses and hemocompatibility of the materials. In vitro experiments showed that the g-HA/PLA composites were well tolerated by the L-929 cells. Hemolysis of the composites was lower than that of pure PLA. Subcutaneous implantation demonstrated that the g-HA/PLA composites were more favorable than the control materials for soft tissue responses. The results suggested that the g-HA/PLA composites are promising and safe materials with potential applications in tissue engineering.


Neonatal hippocampal lesion alters the functional maturation of the prefrontal cortex and the early cognitive development in pre-juvenile rats.
Mnemonic and executive performance is encoded into activity patterns of complex neuronal networks. Lesion studies revealed that adult recognition memory critically depends on the activation of the prefrontal cortex (PFC) and hippocampus (HP). However, its developmental profile remains poorly elucidated. We previously showed the rat PFC and HP are functionally coupled in theta- and gamma-band oscillations during neonatal [postnatal day (P) 5-8] and pre-juvenile (P10-15) stages of development. Here, we assess the behavioral readout of this early prefrontal-hippocampal activation by investigating the ontogeny and the mechanisms of novelty detection and recognition memory in relationship to the functional integrity of the PFC and HP. Excitotoxic lesion of the HP at birth led to abnormal oscillatory entrainment of the PFC throughout neonatal and pre-juvenile development. Although the onset of novelty detection correlated rather with the maturation of sensory perception and motor skills than with hippocampal integrity, the pre-juvenile performance in item, spatial and temporal order recognition memory significantly decreased after HP lesion at birth. This poorer performance does result neither from abnormal developmental milestones and locomotion nor from increased anxiety. Thus, novelty recognition in rat emerges during the second postnatal week and requires functional integrity of communication within neuronal networks including the PFC and HP.


Prespacers formed during primed adaptation associate with the Cas1-Cas2 adaptation complex and the Cas3 interference nuclease-helicase.
For Type I CRISPR-Cas systems, a mode of CRISPR adaptation named priming has been described. Priming allows specific and highly efficient acquisition of new spacers from DNA recognized (primed) by the Cascade-crRNA (CRISPR RNA) effector complex. Recognition of the priming protospacer by Cascade-crRNA serves as a signal for engaging the Cas3 nuclease-helicase required for both interference and primed adaptation, suggesting the existence of a primed adaptation complex (PAC) containing the Cas1-Cas2 adaptation integrase and Cas3. To detect this complex in vivo, we here performed chromatin immunoprecipitation with Cas3-specific and Cas1-specific antibodies using cells undergoing primed adaptation. We found that prespacers are bound by both Cas1 (presumably, as part of the Cas1-Cas2 integrase) and Cas3, implying direct physical association of the interference and adaptation machineries as part of PAC.


Heated hand vein blood is satisfactory for measurements during free fatty acid kinetic studies.
Quantitatively accurate measurements of plasma free fatty acid (FFA) turnover can be made if the FFA tracer is infused intravenously and arterial blood is sampled. To be certain that arterialized-venous blood provides comparable information, plasma FFA concentration and specific activity were measured in both arterial and arterialized-venous blood in 19 volunteers undergoing infusion of [1-14C]palmitate infusion before and during a euglycemic, hyperinsulinemic clamp. Heated hand vein blood was obtained using either the heated box (n = 9 subjects) or heating pad (n = 10 subjects) technique. The heated box arterialized-venous blood had palmitate concentration and specific activity equal to arterial blood. The use of the heating pad resulted in small (3%), but statistically significant (P less than .05), underestimates of palmitate specific activity. In addition, plasma lactate concentrations were significantly higher in heated hand vein blood than arterial blood. In summary, heated hand vein blood is a satisfactory alternative to arterial blood for tracer studies of FFA metabolism.


[Overweight and obesity in children treated for congenital heart disease].
The negative impact of overweight and obesity is potentially greater in children affected by a congenital heart disease (CHD). The aim of this study is to calculate the proportion of overweight and obesity in children who underwent an intervention for CHD, and to investigate systolic arterial hypertension as a possible early cardiovascular complication. A retrospective study was conducted on patients aged 6-17 years treated for CHD, and healthy control subjects, followed-up in a Paediatric Cardiology Clinic. Body mass index percentiles were calculated according to the criteria of WHO. A review was performed on the anthropometric and clinical data, as well as the systolic blood pressure (SBP). A total of 440 patients were included, of which 220 had CHD. The proportion of combined obesity and overweight (body mass index percentile ≥85) was 36.4% (37.3% in healthy subjects and 35.4% in patients with CHD, P=.738). A higher prevalence of obesity (body mass index percentile ≥97) was found in CHD patients (22.7%) compared to 15.5% in healthy subjects (P=.015). SBP percentiles were higher in overweight compared to normal-weight patients (P < .001). The prevalence of SBP readings ≥ the 95th percentile was greater in overweight than in normal weight CHD patients (29.5% versus 7.7%, P < .001) and also in the overweight healthy controls compared to those of normal weight (12.2% versus 0.7%, P < .001). The proportion of obesity is high in children treated for CHD and it is associated with elevated SBP levels. The risk of long-term complications needs to be reduced by means of prevention and treatment of obesity in this vulnerable population.


Towards the development of a fossil bone geochemical standard: an inter-laboratory study.
Ten international laboratories participated in an inter-laboratory comparison of a fossil bone composite with the objective of producing a matrix and structure-matched reference material for studies of the bio-mineralization of ancient fossil bone. We report the major and trace element compositions of the fossil bone composite, using in-situ method as well as various wet chemical digestion techniques. For major element concentrations, the intra-laboratory analytical precision (%RSD(r)) ranges from 7 to 18%, with higher percentages for Ti and K. The %RSD(r) are smaller than the inter-laboratory analytical precision (%RSD(R); <15-30%). Trace element concentrations vary by approximately 5 orders of magnitude (0.1 mg kg(-1) for Th to 10,000 mg kg(-1) for Ba). The intra-laboratory analytical precision %RSD(r) varies between 8 and 45%. The reproducibility values (%RSD(R)) range from 13 to <50%, although extreme value >100% was found for the high field strength elements (Hf, Th, Zr, Nb). The rare earth element (REE) concentrations, which vary over 3 orders of magnitude, have %RSD(r) and %RSD(R) values at 8-15% and 20-32%, respectively. However, the REE patterns (which are very important for paleo-environmental, taphonomic and paleo-oceanographic analyses) are much more consistent. These data suggest that the complex and unpredictable nature of the mineralogical and chemical composition of fossil bone makes it difficult to set-up and calibrate analytical instruments using conventional standards, and may result in non-spectral matrix effects. We propose an analytical protocol that can be employed in future inter-laboratory studies to produce a certified fossil bone geochemical standard.


Evaluation of tetrahydropyranyladriamycin as an immunosuppressant in concordant xenotransplantation.
Tetrahydropyranyladriamycin (THP), a derivative of anthracycline, exhibits a stronger antiproliferative activity and a lower cardiotoxicity than the other anthracyclines. The value of THP as an immunosuppressant has not been examined yet, whereas cyclophosphamide, which is another antiproliferative drug, has shown promise as an immunosuppressant in concordant xenotransplantation. The effect of THP on the marginal zone in the spleen and anti-species antibody production was evaluated and was compared with that of cyclophosphamide in a concordant xenogeneic splenocyte injection model (hamster-to-rat). Next, THP was used as monotherapy for hamster-to-rat heart transplantation to substantiate the potency of its immunosuppressive activity in concordant xenotransplantation. Finally, combination therapy with THP plus FK506 was tried. THP (5 mg/kg) suppressed the expansion of the marginal zone more than did cyclophosphamide (40 mg/kg) and inhibited the production of antispecies antibody as much as did cyclophosphamide (40 mg/kg) in the splenocyte injection model. THP monotherapy could prolong the survival of hamster-to-rat heart grafts up to 14.5 +/- 3.2 days (n = 21). The rejected grafts in the THP-treated animals did not show any histologic evidence of vascular endothelial damage but did exhibit a mononuclear cell infiltration. Combination therapy with THP plus FK506 provided excellent graft survival. THP can prevent early phase humoral rejection in concordant xenotransplantation.


Radioimmunoassay of the anti-cancer agent 4-hydroxyandrostenedione in body fluids.
Antibodies were produced in sheep against a new anti-breast cancer drug 4-hydroxyandrostenedione (4-OHA) using two hapten-ovalbumin conjugates. One of these conjugates (4-hydroxytestosterone-ovalbumin) produced an antiserum suitable for the development of a radioimmunoassay that would allow direct measurement of 4-OHA in plasma at concentrations down to 82 pmol/l, with adequate accuracy, precision and scope for further sensitivity. Although this assay would measure 4-hydroxytestosterone (4-OHT) in addition to 4-OHA, the present data suggest that the magnitude of any interference from endogenous steroids and those derived from 4-OHA could only be minimal. A comparison of solvent-extracted and unextracted samples showed that only unconjugated drug was analysed by this radioimmunoassay. A study of plasma protein binding of 4-OHA showed that at therapeutic concentrations, between 13.5 and 16.5% of plasma 4-OHA was not bound to proteins. This assay system could be a useful adjunct to the future development of 4-OHA as an anti-cancer drug.