Frataxin promotes antioxidant defense in a thiol-dependent manner resulting in diminished malignant transformation in vitro.
Friedreich ataxia is an inherited disorder caused by decreased expression of frataxin protein. Increasing evidence suggests that this protein might detoxify reactive oxygen species (ROS) by an unknown mechanism. Here we demonstrate that transgenic overexpression of human frataxin increases cellular antioxidant defense via activation of glutathione peroxidase and elevation of reduced thiols, thereby reducing the incidence of malignant transformation induced by ROS, as observed by soft agar assays and tumour formation in nude mice. These findings expand the understanding of antioxidant properties of frataxin, and tentatively suggest a role in the early induction of cancer.


Androgen receptor drives transcription of rat PACAP in gonadotrope cells.
Gonadotropin expression is precisely regulated within the hypothalamic-pituitary-gonadal axis through the complex interaction of neuropeptides, gonadal steroids. and both gonadal- and pituitary-derived peptides. In the anterior pituitary gland, the neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) modulates gonadotropin biosynthesis and secretion, acting both alone and in conjunction with GnRH. Steroid hormone feedback also influences gonadotropin expression via both direct and indirect mechanisms. Evidence from nonpituitary tissues suggests that PACAP may be a target for gonadal steroid regulation. In the present study, we show that androgen markedly stimulates rat (r) PACAP promoter-reporter activity in the LβT2 mature mouse gonadotrope cell line. 5'-Serial deletion analysis of reporter constructs identifies 2 regions of androgen responsiveness located at (-915 to -818) and (-308 to -242) of the rPACAP promoter. Androgen receptor (AR) binds directly to DNA cis-elements in each of these regions in vitro. Site-directed mutagenesis of 3 conserved hormone response element half-sites straddling the (-308 to -242) region dramatically blunts androgen-dependent PACAP promoter activity and prevents AR binding at the mutated promoter element. Chromatin immunoprecipitation demonstrates that endogenous AR binds the homologous region on mouse chromatin in LβT2 cells in both the presence and absence of androgen. These data demonstrate that androgen stimulates PACAP gene expression in the pituitary gonadotrope via direct binding of AR to a specific cluster of evolutionarily conserved hormone response elements in the proximal rPACAP gene promoter. Thus, androgen regulation of pituitary PACAP expression may provide an additional layer of control over gonadotropin expression within the hypothalamic-pituitary-gonadal axis.


Ocular histopathology and ultrastructure of Sanfilippo's syndrome, type III-B.
The ocular histopathology of systemic mucopolysaccharidosis, type III-B (Sanfilippo's syndrome) was studied using histochemical and ultrastructural techniques. Cytoplasmic, single-membrane-bound vacuoles containing the major storage product, acid mucopolysaccharide, were found in virtually every ocular tissue. Lamellar cytoplasmic membranous bodies of complex lipid were found mainly in the retinal ganglion cells and the lens epithelium. Many tissues had inclusions that were of an intermediate type and were composed of combined fibrillogranular and lamellar membranous material. Hypopigmentation of the neuroepithelial pigment layers (ie, iris, ciliary, and retinal) seems to be the result of autophagocytosis with melanolysis. Photoreceptor cell degeneration was similar to that seen in some forms of retinitis pigmentosa. The mechanism of photoreceptor cell degeneration is unknown. It may be the result of metabolic dysfunction due to accumulation of mucopolysaccharide in the retinal pigment epithelium.


No Evidence for an Association Between Renal Function and Serious Bleeding Events in Patients Treated With Coumarins: A Population-Based Study.
Although anticoagulation therapy is closely monitored in the Netherlands, coumarin-induced serious bleeding events are still observed. Current literature suggests that renal impairment may contribute to this. To explore the association between renal function and bleeding events during coumarin treatment. A nested case-control study was conducted using data from the PHARMO Database Network. Patients hospitalized for a bleeding event during coumarin treatment were selected as cases and matched on sex, birth year, and geographic region to up to 2 controls using coumarins without hospitalization for bleeding. All values of estimated glomerular filtration rates (eGFRs) were selected in the year before index date (case hospitalization date) and compared between cases and controls using logistic regression analyses. In total, 2224 cases were matched to 4398 controls (61% male; mean ± SD age 75 ± 11 and 78 ± 11 years among cases and controls, respectively). Availability of eGFR values was higher among cases compared with controls (mean ± SD eGFR values 4.5 ± 7.1 vs 3.2 ± 5.5), reflected in the significantly shorter time since last eGFR value (at index date, mean ± SD = 2.7 ± 3.0 vs 3.8 ± 3.1 months; odds ratio [OR] = 0.91, 95%CI = 0.89-0.92). No statistically significant difference was found for the mean eGFR value in the year before index date (mean ± SD 65.7 ± 22.8 vs 64.6 ± 20.9 mL/min/1.73 m2; OR per 10 units [95%CI] = 0.99 [0.96-1.02]). No association between renal function and serious bleeding events during coumarin treatment was observed.


Maternal obesity, pregnancy weight gain, and birth weight and risk of colorectal cancer.
Colorectal cancer (CRC) is a leading cause of cancer-related death worldwide. Obesity is a well-established risk factor for CRC, and fetal or developmental origins of obesity may underlie its effect on cancer in adulthood. We examined associations of maternal obesity, pregnancy weight gain, and birth weight and CRC in adult offspring. The Child Health and Development Studies is a prospective cohort of women receiving prenatal care between 1959 and 1966 in Oakland, California (N=18 751 live births among 14 507 mothers). Clinical information was abstracted from mothers' medical records 6 months prior to pregnancy through delivery. Diagnoses of CRC in adult (age ≥18 years) offspring were ascertained through 2019 by linkage with the California Cancer Registry. We used Cox proportional hazards models to estimate adjusted HR (aHR); we examined effect measure modification using single-referent models to estimate the relative excess risk due to interaction (RERI). 68 offspring were diagnosed with CRC over 738 048 person-years of follow-up, and half (48.5%) were diagnosed younger than age 50 years. Maternal obesity (≥30 kg/m2) increased the risk of CRC in offspring (aHR 2.51, 95% CI 1.05 to 6.02). Total weight gain modified the association of rate of early weight gain (RERI -4.37, 95% CI -9.49 to 0.76), suggesting discordant growth from early to late pregnancy increases risk. There was an elevated association with birth weight (≥4000 g: aHR 1.95, 95% CI 0.8 to 4.38). Our results suggest that in utero events are important risk factors for CRC and may contribute to increasing incidence rates in younger adults.


A comparison of 2-D chromatography separations using UV and (18)O quantification of proteins in similar proteomes.
Proteins present in two mitochondria preparations were separated by 2-D chromatography using the ProteomeLab PF-2D Protein Fractional System, protein fractionation in two dimensions (PF-2D). The proteins in each first-dimension fraction were determined by trypsinization and LC-MS/MS. Chromatography peaks were quantified by UV detection using the "Mapping Tools" software (Beckman). The proteins present in UV detected peaks were trypsinized and identified by automated MS/MS sequencing. Relative amounts of the proteins present in the equivalent peak for each sample were assessed by comparison of the intensities of the constituent peptides and a predicted PF-2D value was calculated from the total ion count (TIC) for each peptide. Relative quantification for (18)O labeled peptides was performed using the ZoomQuant (v1.43b) software [1, 2]. We found that the chromatography peaks detected by UV generally contained several proteins. Using (18)O labeling we determined that in each peak the ratios of the constituent proteins were different. When these ratios were normalized using the TIC to account for abundance, the resulting ratio corresponded to that determined by UV. The predicted value for the PF-2D score corresponded to the observed value for each peak irrespective of the number or proteins detected.


Single atom catalyst-mediated generation of reactive species in water treatment.
Water is one of the most essential components in the sustainable development goals (SDGs) of the United Nations. With worsening global water scarcity, especially in some developing countries, water reuse is gaining increasing acceptance. A key challenge in water treatment by conventional treatment processes is the difficulty of treating low concentrations of pollutants (micromolar to nanomolar) in the presence of much higher levels of inorganic ions and natural organic matter (NOM) in water (or real water matrices). Advanced oxidation processes (AOPs) have emerged as an attractive treatment technology that generates reactive species with high redox potentials (E0) (e.g., hydroxyl radical (HO˙), singlet oxygen (1O2), sulfate radical (SO4˙-), and high-valent metals like iron(IV) (Fe(IV)), copper(III) (Cu(III)), and cobalt(IV) (Co(IV))). The use of single atom catalysts (SACs) in AOPs and water treatment technologies has appeared only recently. This review introduces the application of SACs in the activation of hydrogen peroxide and persulfate to produce reactive species in treatment processes. A significant part of the review is devoted to the mechanistic aspects of traditional AOPs and their comparison with those triggered by SACs. The radical species, SO4˙- and HO˙, which are produced in both traditional and SACs-activated AOPs, have higher redox potentials than non-radical species, 1O2 and high-valent metal species. However, SO4˙- and HO˙ radicals are non-selective and easily affected by components of water while non-radicals resist the impact of such constituents in water. Significantly, SACs with varying coordination environments and structures can be tuned to exclusively generate non-radical species to treat water with a complex matrix. Almost no influence of chloride, carbonate, phosphate, and NOM was observed on the performance of SACs in treating pollutants in water when nonradical species dominate. Therefore, the appropriately designed SACs represent game-changers in purifying water vs. AOPs with high efficiency and minimal interference from constituents of polluted water to meet the goals of water sustainability.


Discovery of potent and selective c-Met inhibitors for MET-amplified hepatocellular carcinoma treatment.
Hepatocellular carcinoma (HCC) is a prevalent and lethal malignancy worldwide. The MET gene, which encodes receptor tyrosine kinase c-Met, is aberrantly activated in various solid tumors, including non-small cell lung cancer and HCC. In this study, we identified a novel c-Met inhibitor 54 by virtual screening and structural optimization. Compound 54 showed potent c-Met inhibition with an IC50 value of 0.45 ± 0.06 nM. It also exhibited high selectivity among 370 kinases and potent anti-proliferative activity against MET-amplified HCC cells. Moreover, compound 54 displayed significant anti-tumor efficacy in vivo, making it a potential candidate for HCC treatment in future studies.


Chest wall motion in preterm infants using respiratory inductive plethysmography.
Preterm infant tidal breathing may be different from that of healthy full-term infants because of various features of the premature thorax. The purpose of this project was to describe chest wall motion in the preterm infant (gestational age <37 weeks) and compare it with chest wall motion data in a group of healthy, full-term infants. We wanted to use an objective bedside method for assessment with minimal disruption to the infant. The study population consisted of 61 preterm human infants whose mean(+/-sD) postconceptional age at time of study was 35.3+/-2.1 weeks. During the study, the infants were quietly awake in a prone position. Preterm infants had initially been admitted to a level III neonatal intensive care unit for acute management and had been transferred to a step-down area, where they were in stable condition for study. Data were collected with a semiquantitatively calibrated, noninvasive respiratory inductive plethysmograph. Mean(+/-SD) phase angle was significantly greater in preterm infants than in full-term infants (60.6+/-39.8 degrees versus 12.5+/-5.0 degrees, respectively, p < or = 0.0001). The laboured breathing index was significantly greater in preterm infants than in full-term infants (1.35+/-0.35 versus 1.01+/-0.01, respectively, p = 0.001). The ribcage contribution to breathing did not differ significantly between preterm and full-term infants (25.5+/-17.7% versus 36.3+/-14.4%, respectively, p = 0.11). These results indicate a significant increase in the degree of ribcage and abdomen asynchrony in the preterm subjects compared to the full-term infants. Plethysmography provided a time-efficient and objective method of assessing chest wall motion in this fragile population.


[Travel during pregnancy (author's transl)].
Before a long journey, the healthy pregnant woman should be advised by her doctor after a careful examination. Healthy pregnant women may consider traveling to any destination except tropical and subtropical regions, where a danger of infection with malaria or dysentery exists. Likewise she should not select a country where vaccination with living virus is required on entry. Pregnant women with a high risk (hemorrhages, toxemia of pregnancy, threatening premature birth) should be advised unconditionally against a long journey.


No Requirement for Targeted Theophylline Levels for Diuretic Effect of Aminophylline in Critically Ill Children.
To determine the relationship between theophylline trough levels and urine output in critically ill children administered aminophylline as adjunctive diuretic therapy. Retrospective cohort study. The PICU of a tertiary care children's hospital. A mixed population of medical/surgical including postoperative cardiothoracic surgery patients less than 18 years old. Electronic medical records of all PICU patients admitted from July 2010 to June 2015 were reviewed, and patients who received aminophylline as diuretic therapy were identified. Patient cohort data including demographics, daily aminophylline, furosemide and chlorothiazide dosing, theophylline trough levels, fluid intake, urine output and total fluid balance, blood urea nitrogen, and creatinine levels were abstracted. Multivariate analysis based on a generalized estimating equations approach demonstrated that aminophylline administration, when analyzed as a categorical variable, was associated with an increase in urine output and decreased fluid balance. However, aminophylline dosing, when analyzed as a continuous variable, was associated with neither an increase in urine output nor decreased fluid balance. Theophylline trough levels were not correlated with urine output at 24 hours (p = 0.78) and were negatively correlated with urine output at 48 hours (r = 0.078; p < 0.005). Aminophylline administration provided a measure of increased diuresis, regardless of dosage, and theophylline trough levels. Therefore, achieving a prescribed therapeutic trough level may not be necessary for full diuretic effect. Because, as opposed to the diuretic effect, the side effect profile of aminophylline is dose-dependent, low maintenance dosing may optimize the balance between providing adjunctive diuretic effect while minimizing the risk of toxicity.


Plant genome editing with TALEN and CRISPR.
Genome editing promises giant leaps forward in advancing biotechnology, agriculture, and basic research. The process relies on the use of sequence specific nucleases (SSNs) to make DNA double stranded breaks at user defined genomic loci, which are subsequently repaired by two main DNA repair pathways: non-homologous end joining (NHEJ) and homology directed repair (HDR). NHEJ can result in frameshift mutations that often create genetic knockouts. These knockout lines are useful for functional and reverse genetic studies but also have applications in agriculture. HDR has a variety of applications as it can be used for gene replacement, gene stacking, and for creating various fusion proteins. In recent years, transcription activator-like effector nucleases and clustered regularly interspaced palindromic repeats (CRISPR) and CRISPR associated protein 9 or CRISPR from Prevotella and Francisella 1 have emerged as the preferred SSNs for research purposes. Here, we review their applications in plant research, discuss current limitations, and predict future research directions in plant genome editing.


Evaluation of Age and Sex-Related Metabolic Changes in Healthy Subjects: An Italian Brain 18F-FDG PET Study.
18F-fluorodeoxyglucose (18F-FDG) positron-emission-tomography (PET) allows detection of cerebral metabolic alterations in neurological diseases vs. normal aging. We assess age- and sex-related brain metabolic changes in healthy subjects, exploring impact of activity normalization methods. brain scans of Italian Association of Nuclear Medicine normative database (151 subjects, 67 Males, 84 Females, aged 20-84) were selected. Global mean, white matter, and pons activity were explored as normalization reference. We performed voxel-based and ROI analyses using SPM12 and IBM-SPSS software. SPM proved a negative correlation between age and brain glucose metabolism involving frontal lobes, anterior-cingulate and insular cortices bilaterally. Narrower clusters were detected in lateral parietal lobes, precuneus, temporal pole and medial areas bilaterally. Normalizing on pons activity, we found a more significant negative correlation and no positive one. ROIs analysis confirmed SPM results. Moreover, a significant age × sex interaction effect was revealed, with worse metabolic reduction in posterior-cingulate cortices in females than males, especially in post-menopausal age. this study demonstrated an age-related metabolic reduction in frontal lobes and in some parieto-temporal areas more evident in females. Results suggested pons as the most appropriate normalization reference. Knowledge of age- and sex-related cerebral metabolic changes is critical to correctly interpreting brain 18F-FDG PET imaging.


Guidelines on the use of cytoreductive surgery and hyperthermic intraperitoneal chemotherapy in patients with peritoneal surface malignancy arising from colorectal or appendiceal neoplasms.
To meet the needs of patients, Canadian surgical and medical oncology leaders in the treatment of peritoneal surface malignancies (psms), together with patient representatives, formed the Canadian HIPEC Collaborative Group (chicg). The group is dedicated to standardizing and improving the treatment of psm in Canada so that access to treatment and, ultimately, the prognosis of Canadian patients with psm are improved. Patients with resectable psm arising from colorectal or appendiceal neoplasms should be reviewed by a multidisciplinary team including surgeons and medical oncologists with experience in treating patients with psm. Cytoreductive surgery and hyperthermic intraperitoneal chemotherapy should be offered to appropriately selected patients and performed at experienced centres. The aim of this publication is to present guidelines that we recommend be applied across the country for the treatment of psm.


Phalangeal osteotomy versus first metatarsal decompression osteotomy for the surgical treatment of hallux rigidus: a prospective study of age-matched and condition-matched patients.
The surgical options for hallux rigidus in the presence of painful but moderate degenerative metatarsophalangeal joint disease are limited to either joint-destructive or joint-preserving procedures. The following study compared the effectiveness of 2 joint-preservation procedures. Forty-nine patients, with a mean age of 53 years, underwent phalangeal osteotomy and were reviewed at an average 29 months postoperatively. A subsequent group of 59 patients, with a mean age of 51 years, underwent first metatarsal decompression osteotomy and were reviewed at an average 15 months postoperatively. In the phalangeal osteotomy group, 65% of patients were completely satisfied, 24% were satisfied with reservation, and 11% were dissatisfied. Three patients suffered continued metatarsophalangeal joint pain, 3 developed hallux interphalangeal joint pain, and 4 patients developed transfer metatarsalgia. The postoperative decrease from 36 degrees to 35 degrees in mean peak hallux dorsiflexion on walking was not significant. In the first metatarsal decompression osteotomy group, 54% were completely satisfied, 13.5% were satisfied with reservations, and 32% were dissatisfied. Continued metatarsophalangeal joint pain occurred in 2 patients, 18 developed transfer metatarsalgia, and 6 of these patients required lesser metatarsal osteotomy. Peak hallux dorsiflexion during walking increased from 36 degrees to 42 degrees (P < .001). First metatarsal decompression osteotomy will increase joint range of motion but the risk of complication and patient dissatisfaction is less after phalangeal osteotomy. Neither procedure could be considered definitive for hallux rigidus.


Why we should care about movements: Using spatially explicit integrated population models to assess habitat source-sink dynamics.
Assessing the source-sink status of populations and habitats is of major importance for understanding population dynamics and for the management of natural populations. Sources produce a net surplus of individuals (per capita contribution to the metapopulation > 1) and will be the main contributors for self-sustaining populations, whereas sinks produce a deficit (contribution < 1). However, making these types of assessments is generally hindered by the problem of separating mortality from permanent emigration, especially when survival probabilities as well as moved distances are habitat-specific. To address this long-standing issue, we propose a spatial multi-event integrated population model (IPM) that incorporates habitat-specific dispersal distances of individuals. Using information about local movements, this IPM adjusts survival estimates for emigration outside the study area. Analysing 24 years of data on a farmland passerine (the northern wheatear Oenanthe oenanthe), we assessed habitat-specific contributions, and hence the source-sink status and temporal variation of two key breeding habitats, while accounting for habitat- and sex-specific local dispersal distances of juveniles and adults. We then examined the sensitivity of the source-sink analysis by comparing results with and without accounting for these local movements. Estimates of first-year survival, and consequently habitat-specific contributions, were higher when local movement data were included. The consequences from including movement data were sex specific, with contribution shifting from sink to likely source in one habitat for males, and previously noted habitat differences for females disappearing. Assessing the source-sink status of habitats is extremely challenging. We show that our spatial IPM accounting for local movements can reduce biases in estimates of the contribution by different habitats, and thus reduce the overestimation of the occurrence of sink habitats. This approach allows combining all available data on demographic rates and movements, which will allow better assessment of source-sink dynamics and better informed conservation interventions.


First metacarpal osteotomy for trapeziometacarpal osteoarthritis.
We report a long-term follow-up of abduction-extension osteotomy of the first metacarpal, performed for painful trapeziometacarpal osteoarthritis. Of a consecutive series of 50 operations, 41 thumbs (82%) were reviewed at a mean follow-up of 6.8 years. Good or excellent pain relief was achieved in 80%, and 93% considered that surgery had improved hand function, while 82% had normal grip and pinch strength, with restoration of thumb abduction. Metacarpal osteotomy was equally successful in relieving symptoms of those with early (grade 2) and moderate (grade 3) degenerative changes. This simple procedure provides lasting pain relief, corrects adduction contracture and restores grip and pinch strength, giving good results with few complications.


Excision of ultraviolet damage and the effect of irradiation on DNA synthesis in a strain of Bloom's syndrome fibroblasts.
We have studied repair of ultraviolet light (UV)-induced damage in a strain of Bloom's syndrome cells which we have shown to be defective in host cell reactivation of UV-irradiated herpes simplex virus. Excision repair was monitored by following loss of sensitivity of DNA in permeabilized cells to digestion by the Micrococcus luteus UV endonuclease preparation. The Bloom's syndrome fibroblasts apparently removed endonuclease-sensitive sites from the DNA slightly less efficiently than did normal strains. After 24 hr, 38% of the sites remained in the Bloom's syndrome cells in comparison with 16% in normal fibroblasts. DNA newly synthesized in UV-irradiated Bloom's syndrome cells sedimented less far into alkaline sucrose gradients than did DNA from similarly treated normal cells. In other respects, including the effect of caffeine exposure, DNA synthesis in Bloom's syndrome cells was indistinguishable from that in normal cells. We were therefore able to detect only minor defects in the repair of UV-induced damage in Bloom's syndrome fibroblasts. This is consistent with the normal survival exhibited by these cells. The defect in excision repair may, however, be sufficient to allow the cellular repair capacity to become saturated at high infecting multiplicities of UV-irradiated herpes simplex virus.


Guidelines for standard preclinical experiments in the mouse model of myasthenia gravis induced by acetylcholine receptor immunization.
Myasthenia gravis (MG) is an autoimmune disorder characterized by generalized muscle weakness due to neuromuscular junction (NMJ) dysfunction brought by acetylcholine receptor (AChR) antibodies in most cases. Although steroids and other immunosuppressants are effectively used for treatment of MG, these medications often cause severe side effects and a complete remission cannot be obtained in many cases. For pre-clinical evaluation of more effective and less toxic treatment methods for MG, the experimental autoimmune myasthenia gravis (EAMG) induced by Torpedo AChR immunization has become one of the standard animal models. Although numerous compounds have been recently proposed for MG mostly by using the active immunization EAMG model, only a few have been proven to be effective in MG patients. The variability in the experimental design, immunization methods and outcome measurements of pre-clinical EAMG studies make it difficult to interpret the published reports and assess the potential for application to MG patients. In an effort to standardize the active immunization EAMG model, we propose standard procedures for animal care conditions, sampling and randomization of mice, experimental design and outcome measures. Utilization of these standard procedures might improve the power of pre-clinical EAMG experiments and increase the chances for identifying promising novel treatment methods that can be effectively translated into clinical trials for MG.


Addressing Urgent Questions for PFAS in the 21st Century.
Despite decades of research on per- and polyfluoroalkyl substances (PFAS), fundamental obstacles remain to addressing worldwide contamination by these chemicals and their associated impacts on environmental quality and health. Here, we propose six urgent questions relevant to science, technology, and policy that must be tackled to address the "PFAS problem": (1) What are the global production volumes of PFAS, and where are PFAS used? (2) Where are the unknown PFAS hotspots in the environment? (3) How can we make measuring PFAS globally accessible? (4) How can we safely manage PFAS-containing waste? (5) How do we understand and describe the health effects of PFAS exposure? (6) Who pays the costs of PFAS contamination? The importance of each question and barriers to progress are briefly described, and several potential paths forward are proposed. Given the diversity of PFAS and their uses, the extreme persistence of most PFAS, the striking ongoing lack of fundamental information, and the inequity of the health and environmental impacts from PFAS contamination, there is a need for scientific and regulatory communities to work together, with cooperation from PFAS-related industries, to fill in critical data gaps and protect human health and the environment.


Design and In-silico Screening of Peptide Nucleic Acid (PNA) Inspired Novel Pronucleotide Scaffolds Targeting COVID-19.
The outburst of the novel coronavirus COVID-19, at the end of December 2019 has turned into a pandemic, risking many human lives. The causal agent being SARS-CoV-2, a member of the long-known Coronaviridae family, is a positive-sense single-stranded enveloped virus and closely related to SARS-CoV. It has become the need of the hour to understand the pathophysiology of this disease, so that drugs, vaccines, treatment regimens and plausible therapeutic agents can be produced. In this regard, recent studies uncovered the fact that the viral genome of SARS-CoV-2 encodes non-structural proteins like RNA-dependent RNA polymerase (RdRp) which is an important tool for its transcription and replication process. A large number of nucleic acid-based anti-viral drugs are being repurposed for treating COVID-19 targeting RdRp. Few of them are at the advanced stage of clinical trials, including remdesivir. While performing a detailed investigation of the large set of nucleic acid-based drugs, we were surprised to find that the synthetic nucleic acid backbone has been explored very little or rare. We designed scaffolds derived from peptide nucleic acid (PNA) and subjected them to in- -silico screening systematically. These designed molecules have demonstrated excellent binding towards RdRp. Compound 12 was found to possess a similar binding affinity as remdesivir with comparable pharmacokinetics. However, the in-silico toxicity prediction indicates that compound 12 may be a superior molecule which can be explored further due to its excellent safety-profile with LD50 12,000mg/kg as opposed to remdesivir (LD50 =1000mg/kg). Compound 12 falls in the safe category of class 6. Synthetic feasibility, equipotent binding and very low toxicity of this peptide nucleic acid-derived compound can make it a leading scaffold to design, synthesize and evaluate many similar compounds for the treatment of COVID-19.


PMA induces androgen receptor downregulation and cellular apoptosis in prostate cancer cells.
Phorbol 12-myristate 13-acetate (PMA) induces cellular apoptosis in prostate cancer cells, the growth of which is governed by androgen/androgen receptor (AR) signaling, but the mechanism by which PMA exerts this effect remains unknown. Therefore, in this study, we investigated the mechanistic action of PMA in prostate cancer cells with regard to AR. We showed that PMA decreased E2F1 as well as AR expression in androgen-dependent prostate cancer LNCaP cells. Furthermore, PMA activated JNK and p53 signaling, resulting in the induction of cellular apoptosis. In LNCaP cells, androgen deprivation and a novel anti-androgen enzalutamide (MDV3100) augmented cellular apoptosis induced by PMA. Moreover, castration-resistant prostate cancer (CRPC) C4-2 cells were more sensitive to PMA compared with LNCaP cells and were sensitized to PMA by enzalutamide. Finally, the expression of PKC, E2F1, and AR was diminished in PMA-resistant cells, indicating that the gain of independence from PKC, E2F1, and AR functions leads to PMA resistance. In conclusion, PMA exerted its anti-cancer effects via the activation of pro-apoptotic JNK/p53 and inhibition of pro-proliferative E2F1/AR in prostate cancer cells including CRPC cells. The therapeutic effects of PMA were augmented by androgen deletion and enzalutamide in androgen-dependent prostate cancer cells, as well as by enzalutamide in castration-resistant cells. Taken together, PMA derivatives may be promising therapeutic agents for treating prostate cancer patients including CRPC patients.


Reliability of brain-computer interface language sample transcription procedures.
We tested the reliability of transcribing language samples of daily brain-computer interface (BCI) communication recorded as language activity monitoring (LAM) logfiles. This study determined interrater reliability and interjudge agreement for transcription of communication of veterans with amyotrophic lateral sclerosis using a P300-based BCI as an augmentative and alternative communication (AAC) system. KeyLAM software recorded logfiles in a universal logfile format during use of BCI-controlled email and word processing applications. These logfiles were encrypted and sent to our laboratory for decryption, transcription, and analysis. The study reports reliability results on transcription of 345 daily logfile samples. The procedure was found to be accurate across transcribers/raters. Frequency of agreement ratios of 97.6% for total number of words and 93.5% for total utterances were found as measures of interrater reliability. Interjudge agreement was 100% for both measures. The results indicated that transcribing language samples using LAM data is highly reliable and the fidelity of the process can be maintained. LAM data supported the transcription of a large number of samples that could not have been completed using audio and video recordings of AAC speakers. This demonstrated efficiency of LAM tools to measure language performance benefits to BCI research and clinical communities.


Adaptive Filter Model of Cerebellum for Biological Muscle Control With Spike Train Inputs.
Prior applications of the cerebellar adaptive filter model have included a range of tasks within simulated and robotic systems. However, this has been limited to systems driven by continuous signals. Here, the adaptive filter model of the cerebellum is applied to the control of a system driven by spiking inputs by considering the problem of controlling muscle force. The performance of the standard adaptive filter algorithm is compared with the algorithm with a modified learning rule that minimizes inputs and a simple proportional-integral-derivative (PID) controller. Control performance is evaluated in terms of the number of spikes, the accuracy of spike input locations, and the accuracy of muscle force output. Results show that the cerebellar adaptive filter model can be applied without change to the control of systems driven by spiking inputs. The cerebellar algorithm results in good agreement between input spikes and force outputs and significantly improves on a PID controller. Input minimization can be used to reduce the number of spike inputs, but at the expense of a decrease in accuracy of spike input location and force output. This work extends the applications of the cerebellar algorithm and demonstrates the potential of the adaptive filter model to be used to improve functional electrical stimulation muscle control.


Chemical constituents of Trichosanthes kirilowii and their cytotoxic activities.
One new lignan, trichobenzolignan (1), and seven known compounds, ligballinol (2), (-)-pinoresinol (3), ehletianol C (4), luteolin 7-O-β-D-glucopyranoside (5), chrysoeriol-7- O-β-D-glucopyranoside (6), 10α-cucurbita-5,24-dien-3β-ol (7), and arvenin I (8). Their structures were established on the basis of spectral and chemical evidence, which were in agreement with those reported in literature. The cytotoxic activities of these compounds were evaluated on four cancer cell lines such as A-549 (human lung cancer), HT-29 (human colon adenocarcinoma), OVCAR (human ovarian carcinoma), and MCF-7 (human breast cancer). As the results, compound 7 showed significant activity on HT-29 and OVCAR cancer cell lines with IC50 of 4.1 and 6.5 µM, respectively. Compounds 1, 5, 6, and 8 exhibited moderate activities in all cancer cell lines with IC50 ranging from 11.3 to 42.8 µM.


Endoneurial potassium is increased and enhances spontaneous activity in regenerating mammalian nerve fibers--implications for neuropathic positive symptoms.
The endoneurial fluid potassium [K+]e has potent effects on membrane potential and excitability but direct measurements have not been reported. The [K+]e was measured with K+ ion-sensitive microelectrodes in normal, demyelinated, and regenerating rat peripheral nerve fibers. The mean value in normal nerve was 3.7 mM. The [K+]e was consistently elevated in regenerating nerves and infrequently elevated in demyelinated nerves. The time course of [K+]e was studied in the left tibial nerve of 22 rats 2 days to 3 months in the extended crush model. The [K+]e was increased and peaked in the first 2 weeks, with a progressive decline toward control values after that. Single fiber recordings were made from the tibial division of the sciatic nerve in 13 animals. The mean number of single fibers studied per rat was 55.5 (SD 24.6). Spontaneous activity (SA) was present in all regenerating nerves and was maximal in the first 2-4 weeks. Endoneurial K+ was further increased by intraarterial injection of potassium into the saphenous artery. Spontaneous activity was enhanced when [K+]e began to rise; there were further superimposed bursts of SA with the administration of additional boluses. These findings suggest that [K+]e increase is a factor in enhancing SA in damaged nerve fibers.


Modular genetic architecture of floral morphology in Nicotiana: quantitative genetic and comparative phenotypic approaches to floral integration.
Animal-pollinated flowers are complex structures that may require a precise configuration of floral organs for proper function. As such, they represent an excellent system with which we can examine the role of phenotypic integration and modularity in morphological evolution. We use complementary quantitative genetic and comparative phenotypic approaches to examine correlations among floral characters in Nicotiana alata, N. forgetiana and their artificial fourth-generation hybrids. Flowers of both species share basic patterns of genetic and phenotypic correlations characterized by at least two integrated character suites that are relatively independent of each other and are not disrupted by four generations of recombination in hybrids. We conclude that these integrated character suites represent phenotypic modules that are the product of a modular genetic architecture. Intrafloral modularity may have been critical for rapid specialization of these species to different pollinators.


Mosquito avoidance and bed net use in the Greater Accra Region, Ghana.
Qualitative research and cross-sectional survey methods were used in a study conducted in rural and urban areas of the Greater Accra Region, Ghana, to explore people's understanding of the cause of malaria and patterns of mosquito avoidance, in particular bed net ownership and use. The study indicated far higher bed net ownership and use in rural than urban areas, which was related partly to perceived affordability and partly to the different contexts of and reasons for avoiding mosquitoes. Knowledge of an association between mosquitoes and malaria, the most common cause of illness in both areas, was related to residence but not to literacy or formal education, and this knowledge did not predict bed net use. The paper points to the complexity of social and personal factors implicated in behavioural interventions for malaria control, and questions behavioural models that assume a linear relationship between knowledge and practice.


Dietary Inclusion of Black Soldier Fly (Hermetia Illucens) Larvae Meal and Paste Improved Gut Health but Had Minor Effects on Skin Mucus Proteome and Immune Response in Atlantic Salmon (Salmo Salar).
The present study investigated effects of dietary inclusion of black soldier fly larvae (BSFL) (Hermetia illucens) meal and paste on gut health, plasma biochemical parameters, immune response and skin mucus proteome in pre-smolt Atlantic salmon (Salmo salar). The seven-week experiment consisted of seven experimental diets: a control diet based on fishmeal and plant protein (Control-1); three BSFL meal diets, substituting 6.25% (6.25IM), 12.5% (12.5IM) and 25% (25IM) of protein; two BSFL paste diets, substituting 3.7% (3.7IP) and 6.7% (6.7IP) of protein and an extra control diet with 0.88% of formic acid (Control-2). The 6.25IM diet reduced enterocyte steatosis in pyloric caeca, improved distal intestine histology, and reduced IgM in distal intestine. The fish fed 12.5IM diet reduced enterocyte steatosis in pyloric caeca, improved distal intestine histology, had a higher plasma lysozyme content compared to 6.25IM, and tend to increase phagocytic activity in head-kidney macrophages-like cells. On the other hand, 25IM diet improved distal intestine histology, but showed mild-moderate enterocyte steatosis in pyloric caeca, increased IFNγ and reduced IgM in distal intestine. In the case of BSFL paste diets, 3.7IP diet caused mild inflammatory changes in distal intestine, although it reduced enterocyte steatosis in pyloric caeca. The 6.7IP diet reduced enterocyte steatosis in pyloric caeca and improved distal intestine histology. Increasing level of BSFL meal in the diet linearly decreased plasma C-reactive protein, whereas increasing level of BSFL paste linearly increased plasma antioxidant capacity. Dietary inclusion of BSFL meal and paste had minor effects on the expression profile of proteins in skin mucus and no effects on immune markers in splenocytes. BSFL meal showed no negative effect on liver and muscle health as indicated by plasma alanine aminotranseferase, asparate aminotransferase and creatine kinase. The present study showed that replacing conventional protein sources with low to moderate levels of BSFL meal (6.25% and 12.5%) or paste (3.7% and 6.7%) reduced enterocyte steatosis in pyloric caeca, while replacing up to 25% with BSFL meal or 6.7% with paste improved distal intestine histology. Further, dietary inclusion of BSFL meal and paste had minor effects on skin mucus proteome and immune response in Atlantic salmon.


2-heptanone, 2-nonanone and 2-undecanone confer oxidation off-flavor in cow milk storage.
Flavor sensation is one of the most prevalent characteristics of food industries and an important consumer preference regulator of dairy products. So far, many volatile compounds have been identified, and their molecular mechanisms conferring overall flavor formation have been reported extensively. However, little is known about the critical flavor compound of a specific sensory in terms of oxidized off-flavor perception. Therefore, the present study aimed to compare the variation in sensory qualities and volatile flavors in full-fat UHT milk (FFM) and low-fat UHT milk (LFM) samples under different natural storage conditions (0, 4, 18, 25, 30, or 37°C for 15 and 30d) and determine the main component causing flavor deterioration in the FFM and LFM samples using sensory evaluation, electronic nose, and HS-SPME-GC-MS. Additionally, the Pearson correlation between the volatile flavor components and oxidative off-flavors was analyzed and validated by sensory reconstitution studies. Compared with the LFM samples, the FFM samples showed a higher degree of quality deterioration with increased storage temperature. Methyl ketones of odd carbon chains, i.e., 2-heptanone, 2-nonanone, 2-undecanone, 2-tridecanone, and 2-pentadecanone reached a maximum content in the FFM37 samples over 30d storage. The combined results of the Pearson correlation and sensory recombination study indicated that 2-heptanone, 2-nonanone, and 2-undecanone conferred off-flavor perception. Overall, the present study results provide potential target components for detecting and developing high-quality dairy products and lay a foundation for specific sensory flavor compound exploration in the food industry.


Dynamic adhesive force measurements under vertical and horizontal motions of interacting rough surfaces.
An instrument to measure dynamic adhesive forces between interacting rough surfaces has been developed. It consists of four parts, namely, main instrument body, vertical positioning system with both micrometer and nanometer positioning accuracies, horizontal positioning system with nanometer positioning accuracy, and custom-built high-resolution, and high dynamic bandwidth capacitive force transducer. The vertical piezoelectric actuator (PZT) controls the vertical (approaching and retracting) motion of the upper specimen, while the horizontal PZT controls the horizontal (reciprocal) motion of the lower specimen. The force transducer is placed in line with the upper specimen and vertical PZT, and directly measures the adhesive forces with a root-mean-square load resolution of 1.7 microN and a dynamic bandwidth of 1.7 kHz. The newly developed instrument enables reliable measurements of near-contact and contact adhesive forces for microscale devices under different dynamic conditions. Using the developed instrument, dynamic pull-in and pull-off force measurements were performed between an aluminum-titanium-carbide sphere and a 10 nm thick carbon film disk sample. Three different levels of contact force were investigated; where for each contact force level the vertical velocity of the upper sample was varied from 0.074 to 5.922 microms, while the lower sample was stationary. It was found that slower approaching and retracting velocities result in higher pull-in and pull-off forces. The noncontact attractive force was also measured during horizontal movement of the lower sample, and it was found that the periodic movements of the lower disk sample also affect the noncontact surface interactions.


Glycosyl-sn-1,2-dimyristylphosphatidylinositol is covalently linked to Trypanosoma brucei variant surface glycoprotein.
The COOH terminus of the externally disposed variant surface glycoprotein (VSG) of the eukaryotic pathogenic protozoan Trypanosoma brucei strain 427 variant MITat 1.4 (117) is covalently linked to a novel phosphatidylinositol-containing glycolipid. This conclusion is supported by analysis of the products of nitrous acid deamination or Staphylococcus aureus phosphatidylinositol-specific phospholipase C treatment of purified membrane-form VSG. Lysis of trypanosomes is accompanied by release of soluble VSG, catalyzed by activation of an endogenous phospholipase C. The only apparent difference between membrane-form VSG and soluble VSG is the removal of sn-1,2-dimyristylglycerol. The COOH-terminal glycopeptide derived by Pronase digestion of soluble VSG was characterized by chemical modification and digestion with alkaline phosphatase. The results are consistent with the single non-N-acetylated glucosamine residue being the reducing terminus of the oligosaccharide and in a glycosidic linkage to a myo-inositol monophosphate that is probably myo-inositol 1,2-cyclic monophosphate. A partial structure for the VSG COOH-terminal moiety is presented. This structure represents a new type of eukaryotic post-translational protein modification and membrane anchor. We discuss the relevance of this structure to observations that have been made with other eukaryotic membrane proteins.


Apnea of prematurity: current management and nursing implications.
Apnea is a common occurrence in preterm infants. Neonatal nurses must have a clear understanding of the diagnosis, pathophysiology, and management of apnea of prematurity. In addition, there are many implications for the nurse caring for the preterm infant with apnea. Nurses play an essential role in monitoring for apnea and the effectiveness of its treatment.


Epidemiology of enteroaggregative Escherichia coli infections and associated outcomes in the MAL-ED birth cohort.
Enteroaggregative E. coli (EAEC) have been associated with mildly inflammatory diarrhea in outbreaks and in travelers and have been increasingly recognized as enteric pathogens in young children with and without overt diarrhea. We examined the risk factors for EAEC infections and their associations with environmental enteropathy biomarkers and growth outcomes over the first two years of life in eight low-resource settings of the MAL-ED study. EAEC infections were detected by PCR gene probes for aatA and aaiC virulence traits in 27,094 non-diarrheal surveillance stools and 7,692 diarrheal stools from 2,092 children in the MAL-ED birth cohort. We identified risk factors for EAEC and estimated the associations of EAEC with diarrhea, enteropathy biomarker concentrations, and both short-term (one to three months) and long-term (to two years of age) growth. Overall, 9,581 samples (27.5%) were positive for EAEC, and almost all children had at least one detection (94.8%) by two years of age. Exclusive breastfeeding, higher enrollment weight, and macrolide use within the preceding 15 days were protective. Although not associated with diarrhea, EAEC infections were weakly associated with biomarkers of intestinal inflammation and more strongly with reduced length at two years of age (LAZ difference associated with high frequency of EAEC detections: -0.30, 95% CI: -0.44, -0.16). Asymptomatic EAEC infections were common early in life and were associated with linear growth shortfalls. Associations with intestinal inflammation were small in magnitude, but suggest a pathway for the growth impact. Increasing the duration of exclusive breastfeeding may help prevent these potentially inflammatory infections and reduce the long-term impact of early exposure to EAEC.


Comparative DNA analyses in longstanding ulcerative colitis with aneuploidy.
In seven patients with longstanding ulcerative colitis, multiple mucosal biopsies taken at colonoscopy were assessed for DNA content and histological dysplasia. DNA analyses were performed using flow cytometric analyses in biopsies and microspectrophotometry in imprint slides prepared from the biopsies. Abnormal aneuploid DNA content was detected in five of the patients in eight separate locations in the colon. There was good conformity between the two methods in the detection of aneuploidy. Both methods of DNA analysis are considered to be applicable in mucosal biopsies from patients with ulcerative colitis. In two patients with significant low grade dysplasia in association with macroscopical lesions (DALMs), there was a relationship with aneuploid DNA pattern found by flow cytometric or microspectrophotometric analyses. Indefinite changes, probably dysplastic were found in three other patients and two of those also displayed aneuploidy, although only once in the same biopsy location. In five locations aneuploidy was detected without concomitant dysplasia. DNA-aneuploidy seems therefore to appear earlier than dysplasia and may be an early marker of malignant transformation of the mucosa in ulcerative colitis. Prospective trials are needed before the clinical significance of early findings of aneuploidy can be determined.


Climatically controlled reproduction drives interannual growth variability in a temperate tree species.
Climatically controlled allocation to reproduction is a key mechanism by which climate influences tree growth and may explain lagged correlations between climate and growth. We used continent-wide datasets of tree-ring chronologies and annual reproductive effort in Fagus sylvatica from 1901 to 2015 to characterise relationships between climate, reproduction and growth. Results highlight that variable allocation to reproduction is a key factor for growth in this species, and that high reproductive effort ('mast years') is associated with stem growth reduction. Additionally, high reproductive effort is associated with previous summer temperature, creating lagged climate effects on growth. Consequently, understanding growth variability in forest ecosystems requires the incorporation of reproduction, which can be highly variable. Our results suggest that future response of growth dynamics to climate change in this species will be strongly influenced by the response of reproduction.


Cell-free hemoglobin mediated oxidative stress is associated with acute kidney injury and renal replacement therapy in severe falciparum malaria: an observational study.
Intravascular hemolysis is an intrinsic feature of severe malaria pathophysiology but the pathogenic role of cell-free hemoglobin-mediated oxidative stress in severe malaria associated acute kidney injury (AKI) is unknown. As part of a prospective observational study, enrolment plasma cell-free hemoglobin (CFH), lipid peroxidation markers (F2-isoprostanes (F2-IsoPs) and isofurans (IsoFs)), red cell deformability, and serum creatinine were quantified in Bangladeshi patients with severe falciparum malaria (n = 107), uncomplicated malaria (n = 80) and sepsis (n = 28). The relationships between these indices and kidney function and clinical outcomes were examined. AKI was diagnosed at enrolment in 58% (62/107) of consecutive patients with severe malaria, defined by an increase in creatinine ≥1.5 times expected baseline. Severe malaria patients with AKI had significantly higher plasma cell-free hemoglobin (geometric mean CFH: 8.8 μM; 95% CI, 6.2-12.3 μM), F2-isoprostane (56.7 pg/ml; 95% CI, 45.3-71.0 pg/ml) and isofuran (109.2 pg/ml; 95% CI, 85.1-140.1 pg/ml) concentrations on enrolment compared to those without AKI (CFH: 5.1 μM; 95% CI, 4.0-6.6 μM; P = 0.018; F2-IsoPs: 27.8 pg/ml; 95% CI, 23.7-32.7 pg/ml; P < 0.001; IsoFs: 41.7 pg/ml; 95% CI, 30.2-57.6 pg/ml; P < 0.001). Cell-free hemoglobin correlated with markers of hemolysis, parasite burden (P. falciparum histidine rich protein 2 (PfHRP2)), and F2-IsoPs. Plasma F2-IsoPs and IsoFs inversely correlated with pH, positively correlated with creatinine, PfHRP2 and fractional excretion of sodium, and were higher in patients later requiring hemodialysis. Plasma F2-IsoP concentrations also inversely correlated with red cell deformability and were higher in fatal cases. Mixed effects modeling including an interaction term for CFH and time showed that F2-IsoPs, IsoFs, PfHRP2, CFH, and red cell rigidity were independently associated with increasing creatinine over 72 h. Multivariable logistic regression showed that admission F2-IsoPs, IsoFs and red cell deformability were associated with the need for subsequent hemodialysis. Cell-free hemoglobin and lipid peroxidation are associated with acute kidney injury and disease severity in falciparum malaria, suggesting a pathophysiological role in renal tubular injury. Evaluation of adjunctive therapies targeting cell-free hemoglobin-mediated oxidative stress is warranted.


Tuning the Seebeck coefficient of naphthalenediimide by electrochemical gating and doping.
We investigate the sign and magnitude of the single-molecule Seebeck coefficient of naphthalenediimide (NDI) under the influence of electrochemical gating and doping. The molecule consists of a NDI core with two alkyl chains in the bay-area position, connected to gold electrodes via benzothiophene (DBT) anchor groups. By switching between the neutral, radical and di-anion charge states, we are able to tune the molecular energy levels relative to the Fermi energy of the electrodes. The resulting single-molecule room-temperature Seebeck coefficents of the three charge states are -294.5 μV K-1, 122 μV K-1 and 144 μV K-1 respectively and the room-temperature power factors are 4.4 × 10-5 W m-1 K-2, 3 × 10-5 W m-1 K-2 and 8.2 × 10-4 W m-1 K-2. As a further strategy for optimising thermoelectric properties, we also investigate the effect on both phonon and electron transport of doping the NDI with either an electron donor (TTF) or an electron acceptor (TCNE). We find that doping by TTF increases the room-temperature Seebeck coefficient and power factor from -73.7 μV K-1 and 2.6 × 10-7 W m-1 K-2 for bare NDI to -105 μV K-1 and 3.6 × 10-4 W m-1 K-2 in presence of TTF. The low thermal conductance of NDI-TTF, combined with the higher Seebeck coefficient and higher electrical conductance lead to a maximum thermoelectric figure of merit of ZT = 1.2, which is higher than that of bare NDI in several orders of magnitude. This demonstrates that both the sign and magnitude of NDI Seebeck coefficient can be tuned reversibly by electrochemical gating and doping, suggesting that such redox active molecules are attractive materials for ultra-thin-film thermoelectric devices.


Mechanisms of trophectoderm fate specification in preimplantation mouse development.
During preimplantation mouse development, embryos establish two distinct cell lineages by the time of blastocyst formation: trophectoderm (TE) and inner cell mass (ICM). To explain the mechanism of this cell fate specification, two classical models, namely the inside-outside model and polarity model have been proposed based on experimental manipulation studies on embryos. This review summarizes recent findings on the molecular mechanisms of fate specification, and discusses how these findings fit into the classical models. TE development is regulated by a transcription factor cascade, the core transcription factors of which are Tead4 and Cdx2. The transcriptional activity of Tead4 is regulated by the position-dependent Hippo signaling pathway, thus supporting the inside-outside model. In contrast, several findings support the polarity model; some other findings suggest different mechanisms. We also discuss how the two classical models could be further developed in the light of recent molecular findings.


In vitro pancreatic hormonal pulses are less regular and more frequent than in vivo.
Spontaneous in vivo cyclic secretion of insulin and glucagon displays a pulse interval of 10 +/- 0.3 (SE) min and a constant phase relationship in fasting rhesus monkeys. When pancreata from six normal rhesus monkeys were perfused in vitro, the insulin pulse interval averaged 6.3 +/- 0.23 (SE) min. Insulin, glucagon, and somatostatin displayed high-amplitude secretory pulses, and the average pulse interval did not differ among the three islet hormones. The islet pulses are less regular in vitro than in vivo, and the phase relationship among the three hormones is lost. The relative amplitude averaged 142 +/- 10, 110 +/- 18, and 81 +/- 11% of the mean hormone concentrations for insulin, somatostatin, and glucagon, respectively. Similar differences in secretory pattern were observed during perfusion of three baboon pancreata compared with the in vivo pattern in this second primate species. The data suggest that the frequency and phase relationship of the islet pulsatile secretory system is modulated by factors extrinsic to the pancreas in the intact nonhuman primate. The nature of these modulating factors remains to be established. The apparent phase independence of the three islet hormones suggests that each of the major endocrine cell types of the islet possess independent episodic secretory mechanisms.


Adverse Childhood Experiences and Arrest Patterns in a Sample of Sexual Offenders.
Developmental psychopathology theories suggest that childhood adversity can contribute to antisocial conduct and delinquent activities. The purpose of this study was to explore the influence of adverse childhood experiences (ACE) on arrest patterns in a sample of sexual offenders (N = 740). Higher ACE scores were associated with a variety of arrest outcomes, indicating that the accumulation of early trauma increased the likelihood of versatility and persistence of criminal behavior. Rapists of adults had higher ACE scores, lower levels of specialization, and higher levels of persistence than sex offenders with minor victims only. Child sexual abuse, emotional neglect, and domestic violence in the childhood home were significant predictors of a higher number of sex crime arrests. For measures of nonsexual arrests and criminal versatility, it was the household dysfunction factors-substance abuse, unmarried parents, and incarceration of a family member-that were predictive, suggesting that family dysfunction and a chaotic home environment contributed significantly to increased risk of general criminal behavior. Sex offenders inspire little sympathy in our society but may be among those most in need of trauma-informed models of treatment that recognize the influence of early adversity on maladaptive schema and self-regulation deficits related to criminal behavior.


A Novel PORCN Frameshift Mutation Leading to Focal Dermal Hypoplasia: A Case Report.
Focal dermal hypoplasia (FDH), also known as Goltz-Gorlin syndrome, is a rare, multisystemic, X-linked dominant genodermatosis characterized by defective development of mesodermal and ectodermal tissues. Major clinical features of the disorder are skin manifestations, skeletal defects, and developmental eye abnormalities. FDH is caused by heterozygous mutations in the PORCN gene located at Xp11.23, and 90% of individuals with FDH are females. Here, we report a female patient with cutaneous changes, multiple eye anomalies, short stature, and ectrodactyly of the right foot. These clinical findings were compatible with the diagnosis of FDH, and a novel mutation, NM_022825.3:c.488delG was found in the PORCN gene causing a premature stop codon.


Professional help-seeking for gambling problems: awareness, barriers and motivators for treatment.
Despite the negative consequences associated with gambling, few problem gamblers seek professional help. This study aimed to examine awareness of professional sources of help and help-seeking behaviour amongst regular and problem gamblers. Australian gamblers (N = 730) were recruited from the general population, multicultural gambling venues, and gambling helplines and treatment services. Surveys measured awareness of professional help services, help-seeking behaviour and motivators and barriers to seeking help. Gamblers demonstrated low awareness of professional help services. Problem gamblers born in Australia or who were divorced were more likely to seek help. Problem gamblers who were reluctant to seek help due to a desire solve the problem on their own and feeling ashamed for themselves or their family pride were more likely to have overcome these barriers to seek help. However, significant barriers related to denial of problem severity and concerns about the ability to access low cost services that cater for multicultural populations predicted a lower likelihood of having sought help. Public education should aim to de-mystify the treatment process and educate gamblers about symptoms of problem gambling to reduce shame, stigma, and denial and encourage help-seeking. Ongoing education and promotion of help services is required to increase awareness of the resources available, including targeted promotions to increase awareness of relevant services among specific populations.


Sugar and fat conditioned flavor preferences in C57BL/6J and 129 mice: oral and postoral interactions.
C57BL/6J (B6) mice consume more sugar and fat solutions than do 129 mice in 24-h preference tests. Previous studies have attributed this observation to strain differences in taste responsiveness to these nutrients. We tested the hypothesis that differences in postingestive responsiveness contribute to the strain differences. In experiment 1, B6 and 129 mice were trained to associate consumption of a flavored solution (CS+) with intragastric (IG) infusions of 16% sucrose and a different flavored solution (CS-) with IG water infusions (22 h/day). They were then retrained with new flavors paired with IG infusions of 5.6% soybean oil and water. Although both strains developed preferences for the nutrient-paired CS+ solutions, the B6 mice displayed significantly stronger preferences. The B6 mice consumed more CS+ during training, which may have contributed to their enhanced preference. In a second experiment, training intakes were equated by giving B6 and 129 mice "isosweet" CS solutions prepared with different amounts of sucrose and saccharin. The B6 and 129 mice consumed more of the sugar- or fat-paired CS+ than the water-paired CS- during training. The two strains also displayed equally strong preferences for the CS+ over CS- in choice tests, indicating that they had similar postingestive responsiveness to the sucrose and soybean oil. We propose that B6 mice consume more sugar and fat than 129 mice because their stronger orosensory response stimulates greater intake, which leads to greater stimulation of postingestive nutrient detectors and further enhancement of consumption.


Effects of parenting style on depressive symptoms, anxiety symptoms, and their comorbidity during the COVID-19 pandemic lockdown among 3117 Chinese junior high school students.
Depressive symptoms, anxiety symptoms, and comorbidity increased in junior high school students due to the outbreak of COVID-19. The objective of this study was to measure the impacts of parenting style on depressive symptoms, anxiety symptoms, and their comorbidity in Chinese junior high school students during the COVID-19 pandemic lockdown period. An online survey was conducted in June 2020 among 3117 junior high school students from Shandong Province, China. The Egna Minnen av Barndoms Uppfostran scale was used to measure parenting styles. The 9-item Patient Health Questionnaire scale and the 7-item Generalized Anxiety Disorder scale were used to measure depressive symptoms and anxiety symptoms, respectively. Multinomial logistic regression was conducted with SPSS 25.0 Version. 3117 junior school students enrolled in this study. The prevalence of depressive symptoms only, anxiety symptoms only, and comorbidity among junior high school students was 11.55 %, 5.29 %, and 22.97 %. Paternal over-protection was a risk factor not only for depressive symptoms only (OR = 1.075, 95 % CI = 1.020-1.134) but for anxiety symptoms only (OR = 1.090, 95 % CI = 1.016-1.170) and comorbidity (OR = 1.098, 95 % CI = 1.050-1.148). Paternal over-interference was a protective factor for depressive symptoms only (OR = 0.947, 95 % CI = 0.908-0.987) and comorbidity (OR = 0.953, 95 % CI = 0.921-0.986). However, maternal over-interference and over-protection were risk factors for depressive symptoms only (OR = 1.039, 95 % CI = 1.011-1.068). This was a cross-sectional study and the causal inferences could not be conducted. Junior high school students with poorer family environmental factors were more likely to suffer from the comorbidity of anxiety symptoms and depressive symptoms during the COVID-19 pandemic lockdown. Decision-makers from the government, educational, and healthcare institutions should pay more attention to junior high school students at higher risk of mental disorders due to poor parenting styles. We should discuss family interventions in the future to prevent mental disorders in junior high school students.


Fat and gastro-oesophageal reflux disease.
It is common practice to prescribe avoidance of fatty foods to patients with gastro-oesophageal reflux disease; however, there is no good evidence supporting the benefit of such a prescription. Several reports looking at gastro-oesophageal reflux after meals of different fat content have been controversial probably because of differences in caloric load, volume and osmolarity, factors which may independently influence reflux. Recent observations suggest that fatty meals do not promote gastro-oesophageal reflux nor alter the competence of the oesophagogastric junction in comparison to balanced meals, provided they contain the same energy load. The possibility that fat increases heartburn by modulating perception of acid reflux seems to have little relevance. It is concluded that, in the light of present evidence, there is no sound rationale for clinicians recommending that patients with GORD follow a low-fat diet.


Angiotensin-II-induced apoptosis requires regulation of nucleolin and Bcl-xL by SHP-2 in primary lung endothelial cells.
Angiotensin II (Ang II) is a key proapoptotic factor in fibrotic tissue diseases. However, the mechanism of Ang-II-induced cell death in endothelial cells has not been previously elucidated. Using the neutral comet assay and specific receptor antagonists and agonists, we found that Ang-II-mediated apoptosis in primary pulmonary endothelial cells required the AT2 receptor. Ang II caused cytochrome c release from the mitochondria concurrent with caspase-3 activation and DNA fragmentation, and apoptosis was suppressed by an inhibitor of Bax-protein channel formation, implicating mitochondrial-mediated apoptosis. There was no evidence that the extrinsic apoptotic pathway was involved, because caspase-9, but not caspase-8, was activated by Ang-II treatment. Apoptosis required phosphoprotein phosphatase activation, and inhibition of the SHP-2 phosphatase (encoded by Ptpn11) blocked cell death. Reduced levels of anti-apoptotic Bcl-2-family members can initiate intrinsic apoptosis, and we found that Ang-II treatment lowered cytosolic Bcl-x(L) protein levels. Because the protein nucleolin has been demonstrated to bind Bcl-x(L) mRNA and prevent its degradation, we investigated the role of nucleolin in Ang-II-induced loss of Bcl-x(L). RNA-immunoprecipitation experiments revealed that Ang II reduced the binding of nucleolin to Bcl-x(L) mRNA in an AU-rich region implicated in instability of Bcl-x(L) mRNA. Inhibition of SHP-2 prevented Ang-II-induced degradation of Bcl-x(L) mRNA. Taken together, our findings suggest that nucleolin is a primary target of Ang-II signaling, and that Ang-II-activated SHP-2 inhibits nucleolin binding to Bcl-x(L) mRNA, thus affecting the equilibrium between pro- and anti-apoptotic members of the Bcl-2 family.


Overexpression of HER-2 in ovarian carcinomas.
The transmembrane receptor encoded by the HER-2 cellular oncogene is amplified in several types of human carcinomas and provides an attractive therapeutic target. Shown by immunohistology, <25% of newly diagnosed ovarian carcinomas express the HER-2 protein. However, now we report that this protein was expressed in all 20 tumor cell lines derived from stage III and IV ovarian cancers as well as in tumor cells harvested from patients with malignant ascites and in tumor samples taken at a second surgery, suggesting that cells with excess expression may have a selective growth advantage. HER-2-positive ovarian carcinoma cells were shown to be sensitive to antibody-dependent cellular cytotoxicity, and their in vitro proliferation was inhibited by anti-HER-2 MAb Herceptin. We postulate, therefore, that therapy which targets HER-2 may be more efficacious in patients with ovarian carcinoma than indicated by the commonly low expression of HER-2 in tumors removed at the time of primary surgery.


Evaluating the effect of body fluid mixture on the relative expression ratio of blood-specific RNA markers.
The estimation of the time elapsed since a biological stain was deposited at a crime scene can provide crucial information to a forensic investigation, indicating either when a crime was committed, or whether the biological evidence was deposited at the time of a known crime event. This would enable the investigators to limit the number of suspects and to assess alibis. The relative expression ratios (RERs) of body fluid-specific RNA markers are promising molecular tools for indicating the age of biological stains. However, the nature of some forensic samples found at crime scenes could be challenging, as they frequently occur in a mixture of different body fluid types. The research presented here has utilised reverse transcription quantitative PCR (RT-qPCR) to explore the impact of bloodstains being present in mixtures with other body fluids (saliva or semen) on the resulting RERs of blood-specific markers. The expression level of three blood-specific markers (HBA, HBB and miR16) along with two reference genes (18S and U6) were analysed across multiple ageing time points in pure and mixed bloodstains. For some markers, no significant differences were found when comparing RERs in pure and mixed bloodstains, however some RERs were altered in mixed stains. This indicates that the presence of body fluid mixtures may have a significant effect on the RERs of some blood-specific markers. This should therefore be considered when selecting markers for estimating the age of stains, particularly when multiple body fluids are thought to be present.


Methods for improving cancer surveillance data in American Indian and Alaska Native populations.
The misclassification of race decreases the accuracy of cancer incidence data for American Indians and Alaska Natives (AI/ANs) in some central cancer registries. This article describes the data sources and methods that were used to address this misclassification and to produce the cancer statistics used by most of the articles in this supplement. Records from United States cancer registries were linked with Indian Health Service (IHS) records to identify AI/AN cases that were misclassified as non-AI/AN. Data were available from 47 registries that linked their data with IHS, met quality criteria, and agreed to participate. Analyses focused on cases among AI/AN residents in IHS Contract Health Service Delivery Area (CHSDA) counties in 33 states. Cancer incidence and stage data were compiled for non-Hispanic whites (NHWs) and AI/ANs across 6 IHS regions of the United States for 1999 through 2004. Misclassification of AI/AN race as nonnative in central cancer registries ranged from 85 individuals in Alaska (3.4%) to 5297 individuals in the Southern Plains (44.5%). Cancer incidence rates among AI/ANs for all cancers combined were lower than for NHWs, but incidence rates varied by geographic region for AI/ANs. Restricting the rate calculations to CHSDA counties generally resulted in higher rates than those obtained for all counties combined. The classification of race for AI/AN cases in cancer registries can be improved by linking records to the IHS and stratifying by CHSDA counties. Cancer in the AI/AN population is clarified further by describing incidence rates by geographic region. Improved cancer surveillance data for AI/AN communities should aid in the planning, implementation, and evaluation of more effective cancer control and should reduce health disparities in this population.


[Five years experience at a community-based comprehensive geriatric assessment unit in southern Israel].
The health care of the growing number of older people in the population is predominantly provided by the family physician, who is usually faced with time limitations. A community-based Comprehensive Geriatric Assessment (CGA) unit has the potential to provide the family physician with the necessary support required in managing older patients. To describe and review the structure and clinical experience of a community-based CGA unit over a period of five years. The CGA unit comprises a multidisciplinary team that utilizes a comprehensive array of assessment instruments. Specific emphasis is placed on comorbidity, polypharmacy, cognitive and affective status, social support and daily function. Over a period of five years, 456 older patients were evaluated. The mean age was 76.7 +/- 6.1 years and 36.2% were women. A total of 6.57 +/- 2.72 new diagnoses were made, the most common being dementia (60.1%), recurrent falls (43.9%) and social problems (32.5%). There were an average of 7 recommendations provided to the family physician, the majority relating to preventive medicine, fall prevention, improving mobility and social interventions. Based on a sample of files evaluated some six months later, we found that these recommendations had been carried out by family physicians to varying degrees. Prescriptions for new medications were provided for 55.6% of those recommended, 45.5% drugs were discontinued as suggested, and 44.4% investigations were performed. Preventive medicine recommendations were carried out in only 11.4% of patients.


Fontan operation in 176 patients with tricuspid atresia. Results and a proposed new index for patient selection.
Between 1973 and March 1989, 176 patients with tricuspid atresia had the Fontan procedure performed at the Mayo Clinic. Age range at the time of surgery was 7 months to 42 years, with 43 patients (24%) 16 years old or older. Hospital mortality rates were 17% (nine of 54) from 1973 through 1980 and 8% (10 of 122) from 1981 through 1989. There have been 10 late cardiovascular deaths. Postoperative follow-up of 139 survivors (range, 6 months to 14 years; mean, 5.5 years) revealed 91% to be in excellent or good condition and 9% to be in fair or poor condition. Patients in fair or poor condition had poor stamina and/or fluid retention with intermittent pleural effusion, ascites, and so on. Two factors that clearly influence operative and late results are preoperative pulmonary arteriolar resistance (Rpa) and left ventricular diastolic function. A preoperative catheterization index devised by adding Rpa to left ventricular end-diastolic pressure divided by QpI plus QsI may be helpful in selecting candidates most likely to survive and benefit from the Fontan operation. In our experience, if this index is less than 4.0, then the postoperative right atrial mean pressure will be 20 mm Hg or less, a circumstance associated with 95% early and 89% overall survival rates.


The formation of ferritin from apoferritin. Kinetics and mechanism of iron uptake.
Ferritin has a high capacity as an iron store, incorporating some 4500 iron atoms as a microcrystalline ferric oxide hydrate. Starting from apoferritin, or ferritin of low iron content, Fe(2+) and an oxidizing agent, the uptake of iron can be recorded spectrophotometrically. Progress curves were obtained and the reconstituted ferritin was shown by several physical methods to be similar to natural ferritin. The progress curves of iron uptake by apoferritin are sigmoidal; those for ferritins of low iron content are hyperbolic. The rate of iron uptake is dependent on the amount of iron already present in the molecule. The distribution of iron contents among reconstituted ferritin molecules is inhomogeneous. These findings are interpreted in terms of a crystal growth model. The surface area of the crystallites forming inside the protein increases until the molecule is half full, and then declines. This surface controls the rate at which new material is deposited. The experimental results can best be accounted for by a two-stage mechanism, an initial slow ;nucleation' stage, which is apparently zero order with respect to [Fe(2+)], followed by a more rapid ;growth' stage. The rate of Fe(2+) oxidation is increased in the presence of apoferritin as compared with controls. Ferritin can therefore be regarded as an enzyme to which the product remains firmly attached. The protein appears to increase the rate of ;nucleation'. The apparent zero order of this stage suggests the presence of binding sites on the protein, which are saturated with respect to Fe(2+). These sites are presumed also to be oxidation sites. The oxidation and subsequent formation of the ferric oxide hydrate may proceed according to one of three alternative models.


Cellular Immunotherapy for Septic Shock. A Phase I Clinical Trial.
In septic animal models mesenchymal stem (stromal) cells (MSCs) modulate inflammation, enhance tissue repair and pathogen clearance, and reduce death. To conduct a phase I dose escalation trial of MSCs in septic shock with the primary objective of examining the safety and tolerability of MSCs. We enrolled nine participants within 24 hours of admission to the ICU. A control cohort of 21 participants was enrolled before starting the MSC interventional cohort to characterize expected adverse events (AEs) and to serve as a comparator for the intervention cohort. Three separate MSC dose cohorts, with three participants per cohort, received a single intravenous dose of 0.3, 1.0, and 3.0 × 106 cells/kg. A prespecified safety plan monitored participants for the occurrence of AEs; cytokines were collected at prespecified time points. Ages of participants in the interventional versus observational cohorts were median of 71 (range, 38-91) and 61 (range, 23-95). Acute Physiology and Chronic Health Evaluation scores were median of 25 (range, 11-28) and 26 (range, 17-32). MSC doses ranged from 19 to 250 million cells. There were no prespecified MSC infusion-associated or serious unexpected AEs, nor any safety or efficacy signals for the expected AEs or the measured cytokines between the interventional and observational cohorts. The infusion of freshly cultured allogenic bone marrow-derived MSCs, up to a dose of 3 million cells/kg (250 million cells), into participants with septic shock seems safe. Clinical trial registered with www.clinicaltrials.gov (NCT02421484).


Effects of dietary fibers with different physicochemical properties on feeding motivation in adult female pigs.
The satiating effects of dietary fiber may depend more on physicochemical properties of the fiber than on total fiber intake. These properties are expected to affect satiety feelings and feeding motivation due to different effects in the gastrointestinal tract. The aim of the current study was to assess the effects of fibers with varying physicochemical properties (bulkiness, viscosity and fermentability) on feeding motivation in adult female pigs. Sixteen pair-housed pigs received four diets: lignocellulose (LC), pectin (PEC), resistant starch (RS), and control (C) without fiber, in four periods in a Latin square design. Each fiber was fed at a low (L) followed by a high (H) inclusion level (7 days each). At 1h, 3h, and 7h after the morning meal, feeding motivation was assessed in an operant test, where turning a wheel yielded multiple food rewards, and in a runway test, where walking a fixed U-shaped track yielded one food reward. Pigs were observed in their home pen for 6h, using 90-s instantaneous scan sampling. In the operant test, throughout the day feeding motivation was higher for pigs on PEC compared with pigs on LC. In the runway, feeding motivation increased particularly at 1h after the meal for pigs on PEC compared with pigs on RS. Also at 7h, feeding motivation tended to decrease for pigs on RS compared with pigs fed other diets. In their home pen, pigs on PEC showed more feeder-directed behavior compared with pigs on RS. In conclusion, PEC was the least satiating fiber. LC and RS, despite a lower supply of available energy, were the most satiating fibers, possibly due to their bulky and fermentation properties, respectively.


Limitations of the trail making test part-B in assessing frontal executive dysfunction.
Part B of the Trail Making Test (TMT-B) is one of the most widely used neuropsychological tests of "executive" function. A commonly held assumption is that the TMT-B can be used to detect frontal executive dysfunction. However, so far, research evidence has been limited and somewhat inconclusive. In this retrospective study, performance on the TMT-B of 55 patients with known focal frontal lesions, 27 patients with focal non-frontal lesions and 70 healthy controls was compared. Completion time and the number of errors made were examined. Patients with frontal and non-frontal lesions performed significantly worse than healthy controls for both completion time and the number of errors. However, there was no significant difference for both completion time and the number of errors when patients with frontal and non-frontal lesions were compared. Performance was also not significantly different between patients with focal lesions within different regions of the frontal lobe (orbital, left lateral, right lateral, medial). Our findings suggest that the TMT-B is a robust test for detection of brain dysfunction. However, its capacity for detecting frontal executive dysfunction appears rather limited. Clinicians should be cautious when drawing conclusions from performance on the TMT-B alone.


The Lack of Amyloidogenic Activity Is Persistent in Old WT and APPswe/PS1ΔE9 Mouse Retinae.
We have previously reported that vision decline was not associated with amyloidogenesis processing in aging C57BL/6J wild-type (WT) mice and in a mouse model of Alzheimer's disease, the APPswe/PS1ΔE9 transgenic mouse model (APP/PS1). This conclusion was drawn using middle-aged (10-13 months old) mice. Here, we hypothesized that compared with hippocampal and cortical neurons, the weak amyloidogenic activity of retinal neurons may result in a detectable release of amyloid β (Aβ) only in aged mice, i.e., between 14 and 24 months of age. The aim of the present study was thus to follow potential activity changes in the amyloidogenic and nonamyloidogenic pathways of young (4 months) and old (20-24 months) WT and APP/PS1 mice. Our results showed that in spite of retinal activity loss reported by electroretinogram (ERG) recordings, the level of amyloid beta precursor protein (APP) and its derivatives did not significantly vary in the eyes of old vs. young mice. Strikingly, the ectopic expression of human APPswe in APP/PS1 mice did not allow us to detect Aβ monomers at 23 months. In contrast, Aβ was observed in hippocampal and cortical tissues at this age but not at 4 months of life. In contrast, optic nerve transection-induced retinal ganglion cell injury significantly affected the level of retinal APP and the secretion of soluble APP alpha in the vitreous. Collectively, these results suggest that the amyloidogenic and nonamyloidogenic pathways are not involved in visual function decline in aging mice. In WT and APP/PS1 mice, it is proposed that retinal neurons do not have the capacity to secrete Aβ in contrast with other cortical and hippocampal neurons.


Reversible O-O Bond Scission and O2 Evolution at MOF-Supported Tetramanganese Clusters.
The scission of the O-O bond in O2 during respiration and the formation of the O-O bond during photosynthesis are the engines of aerobic life. Likewise, the reduction of O2 and the oxidation of reduced oxygen species to form O2 are indispensable components for emerging renewable technologies, including energy storage and conversion, yet discrete molecule-like systems that promote these fundamental reactions are rare. Herein, we report a square-planar tetramanganese cluster formed by self-assembly within a metal-organic framework that reversibly reduces O2 by four electrons, facilitating the interconversion between molecular O2 and metal-oxo species. The tetranuclear cluster spontaneously cleaves the O-O bond of O2 at room temperature to generate a tetramanganese-bis(μ2-oxo) species, which, in turn, is competent for O-O bond reformation and O2 evolution at elevated temperatures, enabled by the head-to-head orientation of two oxo species. This study demonstrates the viability of four-electron interconversion between molecular O2 and metal-oxo species and highlights the importance of site isolation for achieving multi-electron chemistry at polynuclear metal clusters.


Resistin levels of serum and follicular fluid in non-obese patients with polycystic ovary syndrome during IVF cycles.
To measure serum and follicular resistin, steroids hormone levels in women with PCOS (polycystic ovary syndrome) (BMI (body mass index)<25 kg/m(2)), to assess possible correlations of resistin to hormonal and metabolic parameters and to analyze the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET) in women with PCOS and tubal infertility. We analyzed the clinical outcomes of IVF-ET in women with PCOS (BMI<25 kg/m(2)) and tubal infertility during the years 2002 to 2004 and compared the serum and follicular fluid resistin levels, estradiol (E(2)), progesterone (P), testosterone (T) levels in 20 PCOS and 20 healthy, age-matched women without PCOS during IVF-stimulated cycles. The correlations between the resistin levels and the outcomes of IVF-ET were evaluated. No significant differences in resistin levels of either serum or follicular fluid between PCOS and control group were found. However, resistin levels in serum were higher than that in follicular fluid in both groups. Multiple regression analysis showed that resistin levels in serum did not correlate with BMI, estradiol, LH (luteinizing hormone) and insulin level in fasting blood. No significant correlations were found between follicular fluid reisistin levels and fertilization rate, implantation rate, clinical pregnancy rate or early miscarriage rate in both PCOS and control groups. Our results show that resistin does not have correlation with the hormonal and metabolic parameters as well as the outcomes of IVF. These data suggest that resistin is unlikely to be a local determinant factor in steroidogenesis and growth and maturation of oocytes during IVF-ET in lean women with PCOS.


Acupuncture with manual and electrical stimulation for labour pain: a two month follow up of recollection of pain and birth experience.
In a previous randomised controlled trial we showed that acupuncture with a combination of manual- and electrical stimulation (EA) did not affect the level of pain, as compared with acupuncture with manual stimulation (MA) and standard care (SC), but reduced the need for other forms of pain relief, including epidural analgesia. To dismiss an under-treatment of pain in the trial, we did a long-term follow up on the recollection of labour pain and the birth experience comparing acupuncture with manual stimulation, acupuncture with combined electrical and manual stimulation with standard care. Our hypothesis was that despite the lower frequency of use of other pain relief, women who had received EA would make similar retrospective assessments of labour pain and the birth experience 2 months after birth as women who received standard care (SC) or acupuncture with manual stimulation (MA). Secondary analyses of data collected for a randomised controlled trial conducted at two delivery wards in Sweden. A total of 303 nulliparous women with normal pregnancies were randomised to: 40 min of MA or EA, or SC without acupuncture. Questionnaires were administered the day after partus and 2 months later. Two months postpartum, the mean recalled pain on the visual analogue scale (SC: 70.1, MA: 69.3 and EA: 68.7) did not differ between the groups (SC vs MA: adjusted mean difference 0.8, 95% confidence interval [CI] -6.3 to 7.9 and SC vs EA: mean difference 1.3 CI 95% -5.5 to 8.1). Positive birth experience (SC: 54.3%, MA: 64.6% and EA: 61.0%) did not differ between the groups (SC vs MA: adjusted Odds Ratio [OR] 1.8, CI 95% 0.9 to 3.7 and SC vs EA: OR 1.4 CI 95% 0.7 to 2.6). Despite the lower use of other pain relief, women who received acupuncture with the combination of manual and electrical stimulation during labour made the same retrospective assessments of labour pain and birth experience 2 months postpartum as those who received acupuncture with manual stimulation or standard care. ClinicalTrials.gov: NCT01197950.


[Patient classification systems and nursing fees].
This is the fourth year of five-year research plan sponsored by the National Science Council. The purpose of this study was to use VGH-T Factor Patient Classification Systems to calculate the following nursing Fees: (1) by using time study for each nursing activity, (2) by experts' opinion for ideal nursing activity time and ideal nursing charge to calculate nursing Fees, (3) by using different insurance payments and National Health Insurance payments to compare nursing Fees for four types of patients. The investigation period started on March 14, 1995, and ended on June 30, 1995. Patient Classification Tool was used in four general surgical units. There were 1,028 patients' data for patients' needs used. Reliability and validity were based on inter-rater reliability and experts' opinion. The results showed: (1) based on nursing time, the nursing Fees for Types 1, 2, 3, 4 were NT$314, 649, 895 and 1214; (2) according to experts' opinion, the nursing Fees for Types 1, 2, 3, 4 were NT$377, 786, 1076 and 1463; (3) according to labor insurance payments, the nursing Fees for Types 1, 2, 3, 4 were NT$351, 767, 1235 and 1780; (4) according to National Health Insurance payments, the Fees for the four types were NT$378, 915, 1390 and 1960. There were significant differences between the four types of patients. There were significant differences between the different calculations of nursing Fees. The results help not only in patient classification, but also help insurance companies with reference to payment for nursing fees.


[Pulmonary Artery Transection in Surgery for Coronary Artery Fistula].
Coronary artery fistula is a rare abnormality in the communication between a coronary artery and any of the cardiac chambers or major vessels. At present, there is no standard surgical treatment and the most appropriate method is selected on a case-by-case basis. We report one case of coronary artery fistulae in which pulmonary artery transection was required around the left main trunk (LMT). A 62-year-old man who had coronary artery fistulae with an aneurysm which increased from 12 mm to 16 mm in a two-year span. The fistula was located adjacent to the LMT. A complete aneurysm excision under cardiopulmonary bypass was performed, which required pulmonary artery transection. No postoperative complications occurred. Postoperative coronary computed tomography scan showed intact coronary arteries and complete aneurysm removal.


Exosomes from Nischarin-Expressing Cells Reduce Breast Cancer Cell Motility and Tumor Growth.
Exosomes are small extracellular microvesicles that are secreted by cells when intracellular multivesicular bodies fuse with the plasma membrane. We have previously demonstrated that Nischarin inhibits focal adhesion formation, cell migration, and invasion, leading to reduced activation of focal adhesion kinase. In this study, we propose that the tumor suppressor Nischarin regulates the release of exosomes. When cocultured on exosomes from Nischarin-positive cells, breast cancer cells exhibited reduced survival, migration, adhesion, and spreading. The same cocultures formed xenograft tumors of significantly reduced volume following injection into mice. Exosomes secreted by Nischarin-expressing tumors inhibited tumor growth. Expression of only one allele of Nischarin increased secretion of exosomes, and Rab14 activity modulated exosome secretions and cell growth. Taken together, this study reveals a novel role for Nischarin in preventing cancer cell motility, which contributes to our understanding of exosome biology. SIGNIFICANCE: Regulation of Nischarin-mediated exosome secretion by Rab14 seems to play an important role in controlling tumor growth and migration.See related commentary by McAndrews and Kalluri, p. 2099.


Diabetes causes bone marrow autonomic neuropathy and impairs stem cell mobilization via dysregulated p66Shc and Sirt1.
Diabetes compromises the bone marrow (BM) microenvironment and reduces the number of circulating CD34(+) cells. Diabetic autonomic neuropathy (DAN) may impact the BM, because the sympathetic nervous system is prominently involved in BM stem cell trafficking. We hypothesize that neuropathy of the BM affects stem cell mobilization and vascular recovery after ischemia in patients with diabetes. We report that, in patients, cardiovascular DAN was associated with fewer circulating CD34(+) cells. Experimental diabetes (streptozotocin-induced and ob/ob mice) or chemical sympathectomy in mice resulted in BM autonomic neuropathy, impaired Lin(-)cKit(+)Sca1(+) (LKS) cell and endothelial progenitor cell (EPC; CD34(+)Flk1(+)) mobilization, and vascular recovery after ischemia. DAN increased the expression of the 66-kDa protein from the src homology and collagen homology domain (p66Shc) and reduced the expression of sirtuin 1 (Sirt1) in mice and humans. p66Shc knockout (KO) in diabetic mice prevented DAN in the BM, and rescued defective LKS cell and EPC mobilization. Hematopoietic Sirt1 KO mimicked the diabetic mobilization defect, whereas hematopoietic Sirt1 overexpression in diabetes rescued defective mobilization and vascular repair. Through p66Shc and Sirt1, diabetes and sympathectomy elevated the expression of various adhesion molecules, including CD62L. CD62L KO partially rescued the defective stem/progenitor cell mobilization. In conclusion, autonomic neuropathy in the BM impairs stem cell mobilization in diabetes with dysregulation of the life-span regulators p66Shc and Sirt1.


CFH Y402H polymorphism and response to intravitreal ranibizumab in Brazilian patients with neovascular age-related macular degeneration.
To investigate the association between CFH gene polymorphism and response to ranibizumab in Brazilian patients with neovascular age-related macular degeneration (AMD). 95 patients were genotyped for the CFH rs1061170 (Y402H) single nucleotide polymorphism. Patients with neovascular AMD initially received intravitreal ranibizumab injections for three months and were retreated as needed. Visual acuity (VA) and central retinal thickness (CRT) were measured before treatment and at 1, 3, 6, and 12 months post-treatment. For patients with the TT and TC genotypes, paired comparisons of VA showed a statistically significant improvement when the data obtained at all visits were compared with baseline. Patients homozygous for the risk genotype (CC) did not show a statistically significant improvement when VA obtained at visits 1, 3, 6 and 12 were compared with baseline. For all genotypes, paired comparisons of CRT showed a statistically significant improvement when the data obtained at visits 1, 3, 6 and 12 were compared with baseline. Patients with the CC genotype showed poorer long-term functional response to intravitreal ranibizumab.