Organocatalytic, enantioselective synthesis of benzoxaboroles via Wittig/oxa-Michael reaction Cascade of α-formyl boronic acids.
An unprecedented enantioselective synthesis of 3-substituted benzoxaboroles has been developed. An in situ generated ortho-boronic acid containing chalcone provides the chiral benzoxaboroles via an asymmetric oxa-Michael addition of hydroxyl group attached to the boronic acid triggered by the cinchona alkaloid based chiral amino-squaramide catalysts. In general, good yields with good to excellent enantioselectivities (up to 99%) were obtained. The resulting benzoxaboroles were converted to the corresponding chiral β-hydroxy ketones without affecting the enantioselectivity.


Combined antiangiogenic and immune therapy of prostate cancer.
Experimental studies of antiangiogenic or immune therapy of cancer have generated a great deal of optimism. However, the results of clinical testing of these therapies are below expectations. We hypothesized that the antitumor efficacy can be increased when immune destruction of tumor cell is combined with destruction of tumor vasculature by antiangiogenic drugs. In the present study the therapeutic efficacy of combined antiangiogenic and immune therapy has been tested against the highly aggressive, MHC class I negative murine RM1 prostate tumor. SU6668 was used as the antiangiogenic drug and recombinant murine B7.2-IgG fusion protein was used to stimulate T cell-mediated immune destruction of tumor cells. SU6668 is an inhibitor of the tyrosine kinase activity of three angiogenic receptors VEGFR2 (Flk-1/KDR), PDGFRbeta and FGFR1 that play a crucial role in tumor-induced vascularization. Our studies show that B7.2-IgG treatment of mice with established RM1 prostate tumors resulted in a significant inhibition of tumor growth. Both CD4+ and CD8+ T cells were responsible for this effect. SU6668 therapy substantially inhibited tumor vascularization and tumor growth. When tumor-bearing mice were treated with SU6668 in combination with B7.2-IgG, the antitumor effects were substantially higher than in mice treated separately with SU6668 or B7.2-IgG. Prolonged treatment of mice with SU6668 did not inhibit the immunoreactivity of T lymphocytes. On the contrary, T cells from mice treated with a combination of SU6668 and B7.2-IgG showed higher proliferative responses and cytokine production following anti-CD3 stimulation than T cells of mice treated separately with these modalities. These results indicate that antiangiogenic and immune therapies using SU6668 and B7.2-IgG are compatible and manifest complementary antitumor effects. Combined antiangiogenic and immune therapy might represent a new strategy for cancer treatment.


Disruption of allergenic activity of the major grass pollen allergen Phl p 2 by reassembly as a mosaic protein.
The recognition of conformational epitopes on respiratory allergens by IgE Abs is a key event in allergic inflammation. We report a molecular strategy for the conversion of allergens into vaccines with reduced allergenic activity, which is based on the reassembly of non-IgE-reactive fragments in the form of mosaic proteins. This evolution process is exemplified for timothy grass pollen-derived Phl p 2, a major allergen for more than 200 million allergic patients. In a first step, the allergen was disrupted into peptide fragments lacking IgE reactivity. cDNAs coding for these peptides were reassembled in altered order and expressed as a recombinant mosaic molecule. The mosaic molecule had lost the three-dimensional structure, the IgE reactivity, and allergenic activity of the wild-type allergen, but it induced high levels of allergen-specific IgG Abs upon immunization. These IgG Abs crossreacted with group 2 allergens from other grass species and inhibited allergic patients' IgE binding to the wild-type allergen. The mosaic strategy is a general strategy for the reduction of allergenic activity of protein allergens and can be used to convert harmful allergens into safe vaccines.


The possible involvement of hyperpolarizing mechanisms in histamine-induced relaxation of the rat portal vein.
The present study evaluated the effects of histamine 10(-2) M on longitudinal preparations of rat portal vein. It was observed that histamine 10(-2) M induced relaxation of rat portal vein preparations pre-contracted with phenylephrine 10(-4) M. On the other hand, no pharmacological effects were observed in preparations not pre-contracted. The observed histamine-induced relaxing effect was absent in preparations pre-contracted with KCl (120 mM) or in the presence of depolarizing nutritive solution. However, the histamine-induced relaxation was still present in the endothelium-removed preparations. The histamine-induced relaxation also was not prevented by astemizole (10(-6) M, 10(-5) M and 10(-4) M), cimetidine (10(-5) M, 10(-4) M and 10(-3) M) or thioperamide (10(-6) M, 10(-5) M and 10(-4) M), selective antagonists H(1), H(2) and H(3), respectively. The presence of L-NAME 10(-4) M or L-NAME 10(-4) M plus indomethacin 10(-5) M also did not prevent the histamine-induced relaxation observed in rat portal vein. Thus, the histamine-induced relaxation observed in rat portal vein appears to involve a non-endothelial hyperpolarizing mechanism independent of H(1), H(2) and H(3) receptors.


Breastfeeding and Once-Daily Small-Volume Formula Supplementation to Prevent Infant Growth Impairment.
Randomized controlled trials in Guinea-Bissau and Uganda have revealed that the intensive promotion of exclusive breastfeeding (EBF) impairs growth in early infancy. When newborn growth is impaired, small amounts of formula may be combined with breastfeeding to promote growth. To determine if breastfeeding combined with once-daily formula supplementation improves growth among at-risk newborns, we conducted a pilot randomized controlled trial in Bissau, Guinea-Bissau and Kampala, Uganda. We randomly assigned 324 healthy breastfeeding newborns who weighed 2000 g to 2499 g at birth or <2600 g at 4 days old to once-daily formula feeding through 30 days as a supplement to frequent breastfeeding followed by EBF from 31 days through 6 months, or to EBF through 6 months. The primary outcome was weight-for-age z score (WAZ) at 30 days. Other outcomes included weight-for-length z score (WLZ), length-for-age z score (LAZ), breastfeeding cessation, adverse events, and serious adverse events through 180 days. Daily formula consumption in the intervention group was 31.9 ± 11.8 mL. The random assignment did not impact WAZ, WLZ, LAZ, breastfeeding cessation, adverse events, or serious adverse events through 180 days . In the intervention and control groups, 19 (12%) and 35 (21%) infants, respectively, reported nonformula supplementation in the first 30 days (P = .02). Once-daily formula supplementation for 30 days was well-tolerated, but the small volume consumed did not alter growth through 180 days of age. Further research would be required to determine if larger formula volumes, longer duration of treatment, or more frequent feeding are effective at increasing growth for this at-risk population.


HLA-E and HLA-G expression on porcine endothelial cells inhibit xenoreactive human NK cells through CD94/NKG2-dependent and -independent pathways.
Human NK cells contribute a significant role to host defense as well as xenogeneic cytotoxicity. Previous studies using human 721.221 cell line have shown that peptides derived from the leader sequence of the HLA-G binds and up-regulates the surface expression of HLA-E molecules, which was considered to consequently provide negative signals to human NK cells. However, the direct role of HLA-G in inhibiting human NK cells remains controversial. In this study, we showed that the expression of HLA-G or HLA-E in porcine endothelial cells directly protected sensitive porcine cells from human NK cell-mediated xenogeneic cytotoxicity. Ab blocking assays using F(ab')2 of the HLA class I-specific mAb PA2.6 indicated that the protection was directly mediated by the expression of HLA-G and HLA-E on the porcine cells. The HLA-E-mediated protection was blocked by anti-human CD94 Ab. In addition, the engagement of HLA-E lead to the phosphorylation of the CD94/NKG2 complex and the recruitment of SH2 domain-containing protein phosphatase 1 (SHP-1) to the complex. Therefore, HLA-E protected porcine cells from xenoreactive human NK cells through a CD94/NKG2-dependent pathway. In contrast, HLA-G inhibited human NK cells in the absence of CD94/NKG2 phosphorylation or SHP-1 recruitment, and the inhibition was not blocked by anti-CD94 Ab. Therefore, HLA-G protected porcine cells from human NK cells through a CD94/NKG2-independent pathway. These results demonstrated that both HLA-E and HLA-G could directly inhibit human NK cells in the absence of other endogenous HLA class I molecules. These results also have practical implications in preventing xenograft rejection mediated by human NK cells.


Artificial Intelligence and Human Enhancement: Can AI Technologies Make Us More (Artificially) Intelligent?
This paper discusses two opposing views about the relation between artificial intelligence (AI) and human intelligence: on the one hand, a worry that heavy reliance on AI technologies might make people less intelligent and, on the other, a hope that AI technologies might serve as a form of cognitive enhancement. The worry relates to the notion that if we hand over too many intelligence-requiring tasks to AI technologies, we might end up with fewer opportunities to train our own intelligence. Concerning AI as a potential form of cognitive enhancement, the paper explores two possibilities: (1) AI as extending-and thereby enhancing-people's minds, and (2) AI as enabling people to behave in artificially intelligent ways. That is, using AI technologies might enable people to behave as if they have been cognitively enhanced. The paper considers such enhancements both on the level of individuals and on the level of groups.


Induction of spheroplasts in Capnocytophaga ochracea.
A gentle technique for preparing spheroplasts of Capnocytophaga ochracea strain 25 is described. Cells in the exponential phase were washed with 1.0 M-NaCl, agitated in 1.0 M-NaCl for 2 h at 30 degrees C and exposed to lysozyme in a Tris/salts buffer, pH 7.0. This procedure resulted in 98% spheroplast formation with complete removal of the peptidoglycan layer as detected by both phase-contrast and electron microscopy in combination with chemical analysis.


The use of proteomic biomarkers for improved diagnosis and stratification of schizophrenia patients.
Schizophrenia is characterized by a wide spectrum of clinical manifestations, including strong effects on mood and behavior. Patients can also suffer from serious comorbidities including immune system or metabolic abnormalities. Recent advances using proteomic profiling approaches have increased our understanding of these molecular effects and have laid the groundwork for unraveling the heterogeneity of this broadly defined disease. These findings could lead to improved diagnosis and stratification of patients through identification of biochemically different disease subtypes and personalized medicine approaches. The inclusion of molecular signatures in psychiatry will be an important leap forward in providing more effective treatment of patients suffering from this debilitating disorder.


Evaluating Trends in COVID-19 Research Activity in Early 2020: The Creation and Utilization of a Novel Open-Access Database.
Introduction The coronavirus disease 2019 (COVID-19) pandemic has been unprecedented in recent history. The rapid global spread has demonstrated how the emergence of a novel pathogen necessitates new information to advise both healthcare systems and policy-makers. The directives for the management of COVID-19 have been limited to infection control measures and treatment of patients, which has left physicians and researchers alone to navigate the massive amount of research being published while searching for evidence-based strategies to care for patients. To tackle this barrier, we launched CovidReview.ca, an open-access, continually updated, online platform that screens available COVID-19 research to determine higher quality publications. This paper uses data from this review process to explore the activity and trends of COVID-19 research worldwide over time, while specifically looking at the types of studies being published. Materials and Methods The literature search was conducted on PubMed. Search terms included "COVID-19", "severe acute respiratory syndrome coronavirus 2", "coronavirus 19", "SARS-COV-2", and "2019-nCoV". All articles captured by this strategy were reviewed by a minimum of two reviewers and categorized by type of research, relevant medical specialties, and type of publication. Criteria were developed to allow for inclusion or exclusion to the website. Due to the volume of research, only a level 1 (title and abstract) screen was performed. Results The time period for the analysis was January 17, 2020, to May 10, 2020. The total number of papers captured by the search criteria was 10,685, of which 2,742 were included on the website and 7,943 were excluded. The greatest increase in the types of studies over the 16 weeks was narrative review/expert opinion papers followed by case series/reports. Meta-analyses, systematic reviews, and randomized controlled trials remained the least published types of studies. Conclusions The surge of research that accompanied the COVID-19 pandemic is unparalleled in recent years. From our analysis, it is clear that case reports and narrative reviews were the most widely published, particularly in the earlier days of this pandemic. Continued research that falls higher on the evidence pyramid and is more applicable to clinical settings is warranted.


Remote-controlling chemical reactions by light: towards chemistry with high spatio-temporal resolution.
The foundation of the chemical enterprise has always been the creation of new molecular entities, such as pharmaceuticals or polymeric materials. Over the past decades, this continuing effort of designing compounds with improved properties has been complemented by a strong effort to render their preparation (more) sustainable by implementing atom as well as energy economic strategies. Therefore, synthetic chemistry is typically concerned with making specific bonds and connections in a highly selective and efficient manner. However, to increase the degree of sophistication and expand the scope of our work, we argue that the modern aspiring chemist should in addition be concerned with attaining (better) control over when and where chemical bonds are being made or broken. For this purpose, photoswitchable molecular systems, which allow for external modulation of chemical reactions by light, are being developed and in this review we are covering the current state of the art of this exciting new field. These "remote-controlled synthetic tools" provide a remarkable opportunity to perform chemical transformations with high spatial and temporal resolution and should therefore allow regulating biological processes as well as material and device performance.


Priming-dependent phosphorylation and regulation of the tumor suppressor pVHL by glycogen synthase kinase 3.
Inactivation of the von Hippel-Lindau (VHL) tumor suppressor gene is linked to the development of tumors of the eyes, kidneys, and central nervous system. VHL encodes two gene products, pVHL30 and pVHL19, of which one, pVHL30, associates functionally with microtubules (MTs) to regulate their stability. Here we report that pVHL30 is a novel substrate of glycogen synthase kinase 3 (GSK3) in vitro and in vivo. Phosphorylation of pVHL on serine 68 (S68) by GSK3 requires a priming phosphorylation event at serine 72 (S72) mediated in vitro by casein kinase I. Functional analysis of pVHL species carrying nonphosphorylatable or phosphomimicking mutations at S68 and/or S72 reveals a central role for these phosphorylation events in the regulation of pVHL's MT stabilization (but not binding) activity. Taken together, our results identify pVHL as a novel priming-dependent substrate of GSK3 and suggest a dual-kinase mechanism in the control of pVHL's MT stabilization function. Since GSK3 is a component of multiple signaling pathways that are altered in human cancer, our results further imply that normal operation of the GSK3-pVHL axis may be a critical aspect of pVHL's tumor suppressor mechanism through the regulation of MT dynamics.


Effect of different cleaning regimens on the adhesion of resin to saliva-contaminated ceramics.
The aim of this study was to evaluate the influence of different cleaning regimens on the microshear bond strength (μSBS) of three different all-ceramic surfaces after saliva contamination. Cubic ceramic specimens (3 × 3 × 3 mm(3) ) were prepared from three types of ceramics: zirconium dioxide (Z), leucite-reinforced glass ceramic (E), lithium disilicate glass ceramic (EX; n = 12/subgroup). A total of 144 composite resin cylinders (diameter: 1 mm, height: 3 mm) were prepared. Three human-saliva-contaminated surfaces of ceramic specimens were cleaned with either water spray (WS), with 0.5% sodium hypochlorite solution (HC), or with a cleaning paste (CP). Control surface (C) was not contaminated or cleaned. Composite cylinders were bonded to each surface with a resin luting cement. All specimens were stored at 37°C in deionized water until fracture testing. μSBS tests were performed in a universal testing machine (0.5 mm/min), and the results (MPa ± SD) were statistically analyzed (two-way ANOVA, Bonferroni a = 0.05). Fractured surfaces were analyzed to identify the failure types using an optical microscope at 50× magnification. Two representative specimens from all groups were examined with scanning electron microscopy. μSBS test results were significantly affected by the saliva cleaning regimens (p = 0.01) and the ceramic types (p = 0.03). The interaction terms between the ceramic type and saliva cleaning regimen were also significant (p < 0.05). There were no significant differences among the μSBS values (MPa ± SD) for the Z group (C = 17.5 ± 8.8; WS = 16.0 ± 4.9; HC = 17.6 ± 5.8; CP = 16.6 ± 7.5; p > 0.05). In the EX group, C resulted in significantly higher μSBS values (32.6 ± 7.4) than CP (17.4 ± 8.9), WS (15.6 ± 7.3), and HC (14.3 ± 4.5) (p < 0.05); however, C (20.4 ± 7.1) and HC (19.2 ± 7.5) showed higher μSBS values than CP (13.8 ± 4.8) and WS (10.9 ± 5.7) in the E group. Some cohesive failures within the luting resin were observed in the E and EX groups, whereas only adhesive failures were seen in zirconia groups for all surface treatments. Different ceramic surface cleaning regimens after saliva contamination of the zirconium dioxide revealed μSBS similar to the control group, whereas all surface cleaning regimens tested significantly decreased the bond strength values in the lithium disilicate glass ceramic. The leucite-reinforced glass-ceramic group benefited from 0.5% sodium hypochlorite solution cleaning with increased bond strengths. Adhesive cementation of zirconia presents a clinically challenging protocol, and the cementation surface contamination of the zirconia restorations and the inadequate removal of the contaminants increase the risk of failure, as for all ceramic types. This study demonstrated that surface cleaning regimens should be applied according to different ceramic properties.


[Long-term results of hyperfractionated radiotherapy in patients with stage III-IV oropharyngeal carcinoma].
Radiotherapy was given to 139 patients with oropharyngeal carcinoma (stage III-IV) divided into two groups. Group I included 82 patients who received three daily equal fractions (1 Gy), at 4 hr interval, of a mean total focal dose of 80 Gy. Patients in group II (57) received a mean total dose of 68 Gy in 5 weekly standard fractions. Fractionated treatment was carried out in both groups in three stages at 17-day intervals. Complete remission was achieved in 50 (60.9%) patients in group I and in 31 (54.4%)--group II. Total 5-year and recurrence-free survival was 37.3 +/- 5.4 and 32.4 +/- 5.2% (group I) and 12.5 +/- 4.7 and 10.4 +/- 4.4% in group II, respectively. Radiation injury rates were nearly identical in both groups: 14% for standard radiotherapy and 12% for multiple fractions. The data point to high effectiveness of the latter.


Limited proteolysis of tetanus toxin. Relation to activity and identification of cleavage sites.
Tetanus toxin is synthesized by Clostridium tetani as a 151-kDa peptide chain. The primary gene product is processed post-translationally by removal of the initiating methionine residue, formation of disulfide bridges and limited proteolysis by bacterial or exogenous proteinases. The mature toxins consist of a 52-kDa light chain and a 98-kDa heavy chain, linked together by a disulfide bond. Proteolytic nicking is accompanied by increased pharmacological potency. To identify the structural alterations involved, single-chain toxin has been subjected to limited proteolysis with various enzymes. The new N-termini have been determined by Edman degradation and the C-termini by isolation of short C-terminal peptide fragments and subsequent analysis of the sequence and composition. All two-chain toxins result from proteolytic nicking within the 17-residue segment of residues 445-461. Thus, the protease(s) of the culture broth cleave on the C-terminal side of Glu449 and partially Ala456, giving rise to two heavy chain N-termini. Trypsin and clostripain first attack the C-terminal of Arg454 and later Arg448, whereas endoproteinase Arg-C cleaves the former bond only. Chymotrypsin and endoproteinase Glu-C each split a single peptide bond, i.e. that located after Tyr452 and Glu449, respectively. Papain gives rise to a large number of cleavages within the 17-residue segment, the new C-terminus being Thr445 or Asn446 and the new N-terminus being Asp460 or Leu461. Further papain digestion leads to an additional cleavage within the heavy chain between Ser863 and Lys864. The original N-terminal Pro1 and C-terminal Asp1314, predicted from the nucleotide sequence, are conserved in all proteolytic digests. The pharmacological activity of the various two-chain toxins was 5-11 times that of the single-chain toxin, as estimated from the inhibition of [3H]noradrenaline release from rat-brain homogenate. The present data on the processing and activation by limited proteolysis prove the existence of several active tetanus isotoxins. These data, together with our previous data on the localization of disulfide bridges and sulfhydryl groups (Krieglstein, K., Henschen, A., Weller, U. & Habermann, E. (1990) Eur. J. Biochem. 188, 39-45), provide the detailed protein chemical characterization of the tetanus isotoxins.


Follicular Thyroid Carcinoma in a Developing Country: A 10-Year Retrospective Study.
Background The most common endocrine tumor is thyroid cancer. Follicular thyroid carcinoma (FTC) accounts for 5-10% of all thyroid cancers. Patients with FTC frequently present with more advanced diseases and a higher occurrence of distant metastases because of the propensity of vascular invasion. FTC is mainly treated with surgery while radioactive iodine (RAI) therapy is the main adjuvant therapy according to the American Thyroid Association guidelines. Methodology This was a retrospective observational study of FTC patients aged 18 and above conducted at a tertiary care hospital in Karachi from January 01, 2010 to December 31, 2019. Results A total of 404 patients with thyroid carcinoma were sorted, of which 40 (10.1%) were FTC cases. Overall, 50% of the patients were in the age group of 41-60 years, and the female-to-male ratio was 1.5:1. The majority of patients (60%) presented with neck swelling, followed by bone and lung metastasis in 20% and compressive symptoms in another 20%. On fine needle aspiration cytology (FNAC), 50% had Bethesda category III-IV nodules while 10% had Bethesda category II. Overall, 50% had a total thyroidectomy while 50% had a lobectomy followed by a completion thyroidectomy. On histopathology, 23 (57.5%) patients had minimally invasive FTC while 17 (42.5%) had widely invasive FTC. A total of 17 (42.5%) patients had received RAI 30-100 mCi while 10 (25%) received more than 100 mCi. Conclusions FTC can present with both local or metastatic symptoms. The atypical presentation of metastatic FTC should be considered, diagnosed, and managed early to limit mortality and morbidity. Ultrasound is the best diagnostic investigation of choice followed by FNAC. Surgery is the mainstay of treatment and should be followed by RAI in select cases. Thus, understanding the trend of FTC and proper planning and utilization of the resources will help developing countries in effectively treating the FTC.


Luminal alkalinization attenuates proteinuria-induced oxidative damage in proximal tubular cells.
A highly acidic environment surrounds proximal tubular cells as a result of their reabsorption of HCO(3)(-). It is unclear whether this luminal acidity affects proteinuria-induced progression of tubular cell damage. Here, we investigated the contribution of luminal acidity to superoxide (O(2)(·-)) production induced by oleic acid-bound albumin (OA-Alb) in proximal tubular cells. Acidic media significantly enhanced OA-Alb-induced O(2)(·-) production in the HK-2 proximal tubular cell line. Simultaneous treatment with both OA-Alb and acidic media led to phosphorylation of the intracellular pH sensor Pyk2. Highly phosphorylated Pyk2 associated with activation of Rac1, an essential subcomponent of NAD(P)H oxidase. Furthermore, knockdown of Pyk2 with siRNA attenuated the O(2)(·-) production induced by cotreatment with OA-Alb and acid. To assess whether luminal alkalinization abrogates proteinuria-induced tubular damage, we studied a mouse model of protein-overload nephropathy. NaHCO(3) feeding selectively alkalinized the urine and dramatically attenuated the accumulation of O(2)(·-)-induced DNA damage and proximal tubular injury. Overall, these observations suggest that luminal acidity aggravates proteinuria-induced tubular damage and that modulation of this acidic environment may hold potential as a therapeutic target for proteinuric kidney disease.


Circulating matrix Gla protein is associated with coronary artery calcification and vitamin K status in healthy women.
Matrix Gla protein (MGP) is a vitamin K-dependent protein and an inhibitor of vascular calcification. Vitamin K is required for the carboxylation of MGP and can thereby reduce calcification. Circulating MGP species with different conformations have been investigated as markers for coronary artery calcification (CAC). In high-risk populations, high total uncarboxylated MGP (t-ucMGP) was associated with decreased CAC, while high non-phosphorylated uncarboxylated MGP (dp-ucMGP) was associated with a poor vitamin K status. This cross-sectional study investigated the association of MGP species with CAC, vitamin K status among 200 healthy women. Circulating dp-ucMGP, t-ucMGP and, non-phosphorylated carboxylated MGP (dp-cMGP) levels were measured by ELISA techniques and Agatston score by multi-detector computed tomography. The ratio of uncarboxylated to carboxylated osteocalcin was used as proxy of vitamin K status. A borderline significant (P=.06) association between higher circulating dp-ucMGP levels and high CAC was observed (β=0.091, 95% CI-0.01; 0.19). In the entire study population, high t-ucMGP levels tended to be associated (P=.09) with lower CAC (β=-0.36, 95% CI:-0.78; 0.06). This association strengthened amongst women with CAC to a significant relation between high t-ucMGP levels and lower CAC (β=-0.55, 95% CI-1.01;-0.10). Dp-cMGP was not associated with CAC. Low vitamin K-status was associated with high dp-ucMGP concentrations (β=0.138, 95% CI 0.09; 0.19) but not with other MGP species. These results show that dp-ucMGP may serve as a biomarker of vitamin K status. Circulating dp-ucMGP and t-ucMGP may serve as markers for the extent of CAC, but these findings need to be confirmed.


Otoacoustic Emissions in Rural Nicaragua: Cost Analysis and Implications for Newborn Hearing Screening.
Objective (1) Determine the incidence and risk factors for congenital hearing loss. (2) Perform cost analysis of screening programs. Study Design Proportionally distributed cross-sectional survey. Setting Jinotega, Nicaragua. Subjects and Methods Otoacoustic emissions (OAEs) were used to screen 640 infants <6 months of age from neonatal intensive care unit, institutional, and home birth settings. Data on 15 risk factors were analyzed. Cost of 4 implementation strategies was studied: universal screening, screening at the regional health center (RHC), targeted screening, and screening at the RHC plus targeted screening. Cost-effectiveness analysis over 10 years was based on disability-adjusted life year estimates, with the World Health Organization standard of cost-effectiveness ratio (CER) / gross domestic product (GDP) <3, with GDP set at $4884.15. Results Thirty-eight infants failed the initial OAE (5.94%). In terms of births, 325 (50.8%) were in the RHC, 69 (10.8%) in the neonatal intensive care unit, and 29 (4.5%) at home. Family history and birth defect were significant in univariate analysis; birth defect was significant in multivariate analysis. Cost-effectiveness analysis demonstrated that OAE screening is cost-effective without treatment (CER/GDP = 0.06-2.00) and with treatment (CER/GDP = 0.58-2.52). Conclusions Our rate of OAE failures was comparable to those of developed countries and lower than hearing loss rates noted among Nicaraguan schoolchildren, suggesting acquired or progressive etiology in the latter. Birth defects and familial hearing loss correlated with OAE failure. OAE screening of infants is feasible and cost-effective in rural Nicaragua, although highly influenced by estimated hearing loss severity in identified infants and the high travel costs incurred in a targeted screening strategy.


Novel genes involved in the genetic architecture of temperament in Brahman cattle.
Cattle temperament is a complex and economically relevant trait. The objective of this study was to identify genomic regions and genes associated with cattle temperament. From a Brahman cattle population of 1,370 animals evaluated for temperament traits (Exit velocity-EV, Pen Score-PS, Temperament Score-TS), two groups of temperament-contrasting animals were identified based on their EV-average values ±1/2 standard deviation (SD). To be considered in the calm group, the EV of females ranged between 0.16-1.82 m/s (n = 50) and the EV of males ranged between 0.4-1.56 m/s (n = 48). Females were classified as temperamental if their EV ranged between 3.13-7.66 m/s (n = 46) and males were classified as temperamental if their EV ranged between 3.05-10.83 m/s (n = 45). Selected animals were genotyped using a total of 139,376 SNPs (GGP-HD-150K), evaluated for their association with EV. The Genome-Wide Association analysis (GWAS) identified fourteen SNPs: rs135340276, rs134895560, rs110190635, rs42949831, rs135982573, rs109393235, rs109531929, rs135087545, rs41839733, rs42486577, rs136661522, rs110882543, rs110864071, rs109722627, (P<8.1E-05), nine of them were located on intergenic regions, harboring seventeen genes, of which only ACER3, VRK2, FANCL and SLCO3A1 were considered candidate associated with bovine temperament due to their reported biological functions. Five SNPs were located at introns of the NRXN3, EXOC4, CACNG4 and SLC9A4 genes. The indicated candidate genes are implicated in a wide range of behavioural phenotypes and complex cognitive functions. The association of the fourteen SNPs on bovine temperament traits (EV, PS and TS) was evaluated; all these SNPs were significant for EV; only some were associated with PS and TS. Fourteen SNPs were associated with EV which allowed the identification of twenty-one candidate genes for Brahman temperament. From a functional point of view, the five intronic SNPs identified in this study, are candidates to address control of bovine temperament, further investigation will probe their role in expression of this trait.


ESTRO ACROP guidelines for target volume definition in the treatment of locally advanced non-small cell lung cancer.
Radiotherapy (RT) plays a major role in the curative treatment of locally advanced non-small cell lung cancer (NSCLC). Therefore, the ACROP committee was asked by the ESTRO to provide recommendations on target volume delineation for standard clinical scenarios in definitive (chemo)radiotherapy (RT) and adjuvant RT for locally advanced NSCLC. The guidelines given here are a result of the evaluation of a structured questionnaire followed by a consensus discussion, voting and writing procedure within the committee. Hence, we provide advice for methods and time-points of diagnostics and imaging before the start of treatment planning and for the mandatory and optional imaging to be used for planning itself. Concerning target volumes, recommendations are given for GTV delineation of primary tumour and lymph nodes followed by issues related to the delineation of CTVs for definitive and adjuvant radiotherapy. In the context of PTV delineation, recommendations about the management of geometric uncertainties and target motion are given. We further provide our opinions on normal tissue delineation and organisational and responsibility questions in the process of target volume delineation. This guideline intends to contribute to the standardisation and optimisation of the process of RT treatment planning for clinical practice and prospective studies.


Cell depletion due to diphtheria toxin fragment A after Cre-mediated recombination.
Abnormal cell loss is the common cause of a large number of developmental and degenerative diseases. To model such diseases in transgenic animals, we have developed a line of mice that allows the efficient depletion of virtually any cell type in vivo following somatic Cre-mediated gene recombination. By introducing the diphtheria toxin fragment A (DT-A) gene as a conditional expression construct (floxed lacZ-DT-A) into the ubiquitously expressed ROSA26 locus, we produced a line of mice that would permit cell-specific activation of the toxin gene. Following Cre-mediated recombination under the control of cell-type-specific promoters, lacZ gene expression was efficiently replaced by de novo transcription of the Cre-recombined DT-A gene. We provide proof of this principle, initially for cells of the central nervous system (pyramidal neurons and oligodendrocytes), the immune system (B cells), and liver tissue (hepatocytes), that the conditional expression of DT-A is functional in vivo, resulting in the generation of novel degenerative disease models.


Metallothionein immunohistochemistry has high sensitivity and specificity for detection of Wilson disease.
Diagnosis of Wilson disease (WD) can be difficult because of its protean clinical presentations, but early diagnosis is important because effective treatment is available and can prevent disease progression. Similarly, diagnosis of WD on liver biopsy specimens is difficult due to the wide range of histologic appearances. A stain that could help identify WD patients would be of great value. The goal of this study was to use mass spectrometry-based proteomics to identify potential proteins that are differentially expressed in WD compared to controls, and could serve as potential immunohistochemical markers for screening. Several proteins were differentially expressed in WD and immunohistochemical stains for two (metallothionein (MT) and cytochrome C oxidase copper chaperone (COX17)) were tested and compared to other methods of diagnosis in WD including copper staining and quantitative copper assays. We found diffuse metallothionein immunoreactivity in all liver specimens from patients with WD (n = 20); the intensity of the staining was moderate to strong. This staining pattern was distinct from that seen in specimens from the control groups (none of which showed strong, diffuse staining), which included diseases that may be in the clinical or histologic differential of WD (steatohepatitis (n = 51), chronic viral hepatitis (n = 40), autoimmune hepatitis (n = 50), chronic biliary tract disease (n = 42), and normal liver (n = 20)). COX17 immunostain showed no significant difference in expression between the WD and control groups. MT had higher sensitivity than rhodanine for diagnosis of WD. While the quantitative liver copper assays also had high sensitivity, they require more tissue, have a higher cost, longer turnaround time, and are less widely available than an immunohistochemical stain. We conclude that MT IHC is a sensitive immunohistochemical stain for the diagnosis of WD that could be widely deployed as a screening tool for liver biopsies in which WD is in the clinical or histologic differential diagnosis.


Molecular characterization of murine humoral immune response to botulinum neurotoxin type A binding domain as assessed by using phage antibody libraries.
To produce antibodies capable of neutralizing botulinum neurotoxin type A (BoNT/A), the murine humoral immune response to BoNT/A binding domain (H(C)) was characterized at the molecular level by using phage antibody libraries. Mice were immunized with BoNT/A H(C), the spleens were harvested, and single-chain Fv (scFv) phage antibody libraries were constructed from the immunoglobulin heavy and light chain variable region genes. Phage expressing BoNT/A binding scFv were isolated by selection on immobilized BoNT/A and BoNT/A H(C). Twenty-eight unique BoNT/A H(C) binding scFv were identified by enzyme-linked immunosorbent assay and DNA sequencing. Epitope mapping using surface plasmon resonance in a BIAcore revealed that the 28 scFv bound to only 4 nonoverlapping epitopes with equilibrium constants (Kd) ranging from 7.3 x 10(-8) to 1.1 x 10(-9) M. In a mouse hemidiaphragm assay, scFv binding epitopes 1 and 2 significantly prolonged the time to neuroparalysis, 1.5- and 2.7-fold, respectively, compared to toxin control. scFv binding to epitopes 3 and 4 showed no protection against neuroparalysis. A combination of scFv binding epitopes 1 and 2 had an additive effect on time to neuroparalysis, which increased to 3.9-fold compared to the control. The results suggest that there are two "productive" receptor binding sites on H(C) which lead to toxin internalization and toxicity. Blockade of these two epitopes with monoclonal antibodies may provide effective immunoprophylaxis or therapy against BoNT/A intoxication.


Enhanced visible light promoted antibacterial efficiency of conjugated microporous polymer nanoparticles via molecular doping.
The increase in the resistance of bacteria to antibiotics is one of the main concerns of public health and holds a great demand in the development of new disinfection methods. Photodynamic therapy (PDT) has been considered as a promising alternative approach towards the eradication of bacteria and great attention has been dedicated to the use of non-toxic and pure organic PDT agents. Herein we report the structural design method of a series of conjugated microporous polymer nanoparticles (CMP NPs) as a new class of highly effective photoactive materials for the inactivation of bacteria in water upon visible light exposure. Through molecular doping of electron-withdrawing moieties into electron-donating polymer backbones, enhanced antibacterial properties are demonstrated upon the inactivation of Escherichia coli K-12 and Bacillus subtilis mainly by means of photogeneration of singlet oxygen as the main photogenerated active species. Additionally, the high stability, reusability and disinfection mechanism of the CMP NPs are also described.


Differential bioavailability of polychlorinated biphenyls associated with environmental particles: Microplastic in comparison to wood, coal and biochar.
Microplastic particles are increasingly being discovered in diverse habitats and a host of species are found to ingest them. Since plastics are known to sorb hydrophobic organic contaminants (HOCs) there is a question of what risk of chemical exposure is posed to aquatic biota from microplastic-associated contaminants. We investigate bioavailability of polychlorinated biphenyls (PCBs) from polypropylene microplastic by measuring solid-water distribution coefficients, gut fluid solubilization, and bioaccumulation using sediment invertebrate worms as a test system. Microplastic-associated PCBs are placed in a differential bioavailability framework by comparing the results to several other natural and anthrogenic particles, including wood, coal, and biochar. PCB distribution coefficients for polypropylene were higher than natural organic materials like wood, but in the range of lipids and sediment organic carbon, and smaller than black carbons like coal and biochars. Gut fluid solubilization potential increased in the order: coal < polypropylene < biochar < wood. Interestingly, lower gut fluid solubilization for polypropylene than biochar infers that gut fluid micelles may have solubilized part of the biochar matrix while bioaccessibility from plastic can be limited by the solubilizing potential of gut fluids dependent on the solid to liquid ratio or renewal of fluids in the gut. Biouptake in worms was lower by 76% when PCBs were associated with polypropylene compared to sediment. The presence of microplastics in sediments had an overall impact of reducing bioavailability and transfer of HOCs to sediment-ingesting organisms. Since the vast majority of sediment and suspended particles in the environment are natural organic and inorganic materials, pollutant transfer through particle ingestion will be dominated by these particles and not microplastics. Therefore, these results support the conclusion that in most cases the transfer of organic pollutants to aquatic organisms from microplastic in the diet is likely a small contribution compared to other natural pathways of exposure.


Preview frequency effects in reading: evidence from Chinese.
Studies about sentence reading have shown that visual and lexical information beyond the currently fixated word can be integrated across fixations. The gaze-contingent boundary paradigm has been used widely to explore the extent to which parafoveal information can be processed before a word is fixated on. However, a critical review of the current literature suggests that unrelated mask previews are an unlikely baseline control with zero lexical activation, blurring the nature of experimental effects observed in the paradigm. The present study, therefore, aimed at shedding light on the effect of parafoveal mask properties through a manipulation of preview word frequency. Low-frequency preview words that are unrelated to target words elicited a larger interference than high-frequency preview words. We discuss implications of the preview frequency effect for computational models of eye-movement control in reading.


Age-Adjusted Associations Between Comorbidity and Outcomes of COVID-19: A Review of the Evidence From the Early Stages of the Pandemic.
Objectives: Early in the COVID-19 pandemic, people with underlying comorbidities were overrepresented in hospitalised cases of COVID-19, but the relationship between comorbidity and COVID-19 outcomes was complicated by potential confounding by age. This review therefore sought to characterise the international evidence base available in the early stages of the pandemic on the association between comorbidities and progression to severe disease, critical care, or death, after accounting for age, among hospitalised patients with COVID-19. Methods: We conducted a rapid, comprehensive review of the literature (to 14 May 2020), to assess the international evidence on the age-adjusted association between comorbidities and severe COVID-19 progression or death, among hospitalised COVID-19 patients - the only population for whom studies were available at that time. Results: After screening 1,100 studies, we identified 14 eligible for inclusion. Overall, evidence for obesity and cancer increasing risk of severe disease or death was most consistent. Most studies found that having at least one of obesity, diabetes mellitus, hypertension, heart disease, cancer, or chronic lung disease was significantly associated with worse outcomes following hospitalisation. Associations were more consistent for mortality than other outcomes. Increasing numbers of comorbidities and obesity both showed a dose-response relationship. Quality and reporting were suboptimal in these rapidly conducted studies, and there was a clear need for additional studies using population-based samples. Conclusions: This review summarises the most robust evidence on this topic that was available in the first few months of the pandemic. It was clear at this early stage that COVID-19 would go on to exacerbate existing health inequalities unless actions were taken to reduce pre-existing vulnerabilities and target control measures to protect groups with chronic health conditions.


Mycophenolate mofetil in solid-organ transplantation.
This review focuses on the use of mycophenolate mofetil (MMF) as an immunosuppressive agent in solid-organ transplantation. MMF, a non-competitive inhibitor of inosine monophosphate dehydrogenase, blocks de novo purine synthesis in T and B lymphocytes, resulting in the selective inhibition of proliferation of these cells in response to antigenic stimuli. MMF may also promote apoptosis of these cells. The immunosuppressive ability of MMF is thought to derive mainly from the inhibition of inosine monophosphate dehydrogenase. The other effects of MMF include suppression of antibody synthesis by B lymphocytes, inhibition of proliferation of smooth muscle cells in culture and impaired glycosylation of adhesion molecules. MMF may exhibit anti-inflammatory effects resulting from decreased activity of the inducible form of nitric oxide synthase, a consequence of depletion of tetrahydrobiopterin, which leads to decreased generation of peroxynitrite, a pro-inflammatory molecule. The pharmacokinetics, pharmacodynamics and principles underlying therapeutic drug monitoring of MMF are reviewed. The results of the pivotal clinical trials of MMF in kidney and heart transplantation are discussed and a summary of the major studies demonstrating a positive effect of MMF on renal transplantation outcomes is presented. The use of MMF in the context of ABO-incompatible renal transplantation, renal transplantation in highly sensitised and cross-match positive recipients, humoral rejection of renal allografts, chronic allograft nephropathy and steroid/calcineurin inhibitor minimisation in renal transplantation are also discussed.


Regulation of pyruvate kinase by 6-phosphogluconate in isolated hepatocytes.
Isolated liver parenchymal cells from rats fed a 65% sucrose diet for 14 days were incubated in the presence and absence of 10(-6) M glucagon. The pyruvate kinase obtained from homogenates of the glucagon-treated cells displayed and increased Ks 0.5 for phosphoenolpyruvate (P-enolpyruvate), as well as an increased Ka 0.5 for 6-phosphogluconate (6-P-gluconate), compared to pyruvate kinase from untreated cells. Additionally, glucagon treatment decreased the maximal stimulation of pyruvate kinase by 6-P-gluconate by approximately two-thirds and decreased the Hill coefficient value of pyruvate kinase for 6-P-gluconate from 1.76 to 1.56. 6-Aminonicotinamide, an inhibitor of 6-P-gluconate dehydrogenase, increased 6-P-gluconate levels in isolated liver parenchymal cells three- to sevenfold, depending on the substrates present. The flux of P-enolpyruvate through pyruvate kinase was increased from 18 to 40% in these preparations and was highly correlated with the increase in 6-P-gluconate levels. The results suggest that 6-P-gluconate could regulate pyruvate kinase activity in the intact liver parenchymal cell. Furthermore, the activator would be of greatest importance in the lipogenic animal.


Myocardial perfusion scintigraphy for risk stratification of patients with coronary artery disease: the AMICO registry.
We assessed the prognostic value of myocardial perfusion scintigraphy (MPS) with cadmium-zinc-telluride in addition to clinical and coronary anatomy analysis. We prospectively enrolled 1464 patients (26% females, 69.5 ± 10.4 years) referred for stress-rest MPS. All the patients underwent invasive coronary angiography (1171, 80%) or coronary computed tomography angiography (293, 20%). We defined a composite endpoint of cardiovascular death and non-fatal MI. After an 8-year follow-up, summed stress score (SSS) had the highest accuracy in predicting primary endpoint with a ROC-derived cut-off of SSS >8 (>10% myocardium). SSS >8 portended the lowest survival probability at Kaplan-Meier analysis (P < 0.0001 for the composite endpoint and individual components). The Cox-regression analysis indicated SSS as an independent predictor of the composite endpoint, along with fasting blood glucose and total cholesterol and contrary to coronary anatomy parameters. Patients with SSS >8 treated with optimal medical therapy (OMT) had the largest area of necrosis, the lower ischaemic burden, the most compromised LV systo-diastolic function and the highest LV mass, but received a less aggressive treatment in comparison to early revascularized patients. Survival analysis revealed patients with SSS ≤8 had the greater freedom from events, irrespective of the treatment strategy, while the group with SSS >8 and OMT had the worst outcome, followed by patients with SSS >8 and early revascularization (log-rank test: all P < 0.0001). MPS-SSS constitutes a strong independent predictor of future adverse events after adjustment for multiple clinical parameters and coronary angiography. In particular, MPS could help risk stratification of patients who did not undergo early revascularization.


Mucosal Melanoma Originating From the Eustachian Tube.
A 77-year-old man was referred with a 4-month history of hearing impairment and ear fullness of the left ear. Otoscopic examination revealed an effusion in the left middle ear, and nasal endoscopic examination revealed a dark polypoid lesion at the opening of the left Eustachian tube. In addition to the lesion of the Eustachian tube, a dark mucosal lesion was seen at the posterior choana and the posterior end of the nasal septum. Endoscopic biopsy was done and pathologic result was consistent with malignant melanoma. Wide surgical excision with postoperative radiotherapy was performed; multiple metastases were detected 4 months after the treatment. Mucosal melanoma originating from nasopharynx was extremely rare, but careful examination of nasopharyngeal area should be considered when the patient presents with unilateral middle ear effusion, especially in older age.


Locomotion dynamics of hunting in wild cheetahs.
Although the cheetah is recognised as the fastest land animal, little is known about other aspects of its notable athleticism, particularly when hunting in the wild. Here we describe and use a new tracking collar of our own design, containing a combination of Global Positioning System (GPS) and inertial measurement units, to capture the locomotor dynamics and outcome of 367 predominantly hunting runs of five wild cheetahs in Botswana. A remarkable top speed of 25.9 m s(-1) (58 m.p.h. or 93 km h(-1)) was recorded, but most cheetah hunts involved only moderate speeds. We recorded some of the highest measured values for lateral and forward acceleration, deceleration and body-mass-specific power for any terrestrial mammal. To our knowledge, this is the first detailed locomotor information on the hunting dynamics of a large cursorial predator in its natural habitat.


Effects of vitamin E supplementation on exercise-induced oxidative stress: a meta-analysis.
Tocopherols (commonly referred to as "vitamin E") are frequently studied antioxidants in exercise research. However, the studies are highly heterogeneous, which has resulted in contradicting opinions. The aim of this review is to identify similar studies investigating the effects of tocopherol supplementation on exercise performance and oxidative stress and to perform minimally biased qualitative comparisons and meta-analysis. The literature search and study selection were performed according to Cochrane guidelines. A 2-dimensional study execution process was developed to enable selection of similar and comparable studies. Twenty relevant studies were identified. The high variability of study designs resulted in final selection of 6 maximally relevant studies. Markers of lipid peroxidation (malondialdehyde) and muscle damage (creatine kinase) were the 2 most frequently and similarly measured variables. Meta comparison showed that tocopherol supplementation did not result in significant protection against either exercise-induced lipid peroxidation or muscle damage. The complex antioxidant nature of tocopherols and low accumulation rates in muscle tissues could underlie an absence of protective effects.


Transforming growth factor-beta (TGF-beta).
The TGF-beta family of cytokines are ubiquitous, multifunctional and essential to survival. They play important roles in growth and development, inflammation and repair and host immunity. The mammalian TGF-beta isoforms (TGF-beta 1, beta 2 and beta 3) are secreted as latent precursors and have multiple cell surface receptors of which at least two mediate signal transduction. Autocrine and paracrine effects of TGF-beta's can be modified by extracellular matrix, neighbouring cells and other cytokines. The vital role of the TGF-beta family is illustrated by the fact that approximately 50% of TGF-beta 1 gene knockout mice die in utero and the remainder succumb to uncontrolled inflammation after birth. TGF-beta 2 and TGF-beta 3 gene knockout mice are not yet described. More recently, novel TGF-beta-like molecules have been described which share some of the properties of the mammalian TGF-beta isoforms. The role of TGF-beta in homeostatic and pathogenic processes suggests numerous applications in the diagnosis and treatment of various diseases characterised by inflammation and fibrosis.


HSP90: a rising star on the horizon of anticancer targets.
Over the past decade, heat-shock protein (HSP)90 has begun to draw increasing attention as a novel anticancer target with unique features. As a molecular chaperone, HSP90 promotes the maturation and maintains the stability of a large number of conformationally labile client proteins, most of which are involved in biologic processes that are often deranged within tumor cells, such as signal transduction, cell-cycle progression and apoptosis. As a result, and in contrast to other molecular targeted therapeutics, inhibitors of HSP90 achieve their promising anticancer activity through simultaneous disruption of many oncogenic substrates within cancer cells. This review provides a brief summary of HSP90 biology and its association with cancer. It describes the discovery and development of HSP90 inhibitors as anticancer agents and their current status in the clinic. Finally, it closes with a discussion of the unique challenges confronting the further development of these agents and their prospects for the future.


Droplet spreading driven by van der Waals force: a molecular dynamics study.
The dynamics of droplet spreading is investigated by molecular dynamics simulations for two immiscible fluids of equal density and viscosity. All the molecular interactions are modeled by truncated Lennard-Jones potentials and a long-range van der Waals force is introduced to act on the wetting fluid. By gradually increasing the coupling constant in the attractive van der Waals interaction between the wetting fluid and the substrate, we observe a transition in the initial stage of spreading. There exists a critical value of the coupling constant, above which the spreading is pioneered by a precursor film. In particular, the dynamically determined critical value quantitatively agrees with that determined by the energy criterion that the spreading coefficient equals zero. The latter separates partial wetting from complete wetting. In the regime of complete wetting, the radius of the spreading droplet varies with time as [Formula: see text], a behavior also found in molecular dynamics simulations where the wetting dynamics is driven by the short-range Lennard-Jones interaction between liquid and solid.


Cationic polymers and their self-assembly for antibacterial applications.
The present article focuses on the amphiphilic cationic polymers as antibacterial agents. These polymers undergo self-assembly in aqueous conditions and impart biological activity by efficiently interacting with the bacterial cell wall, hence, used in preparing chemical disinfectants and biocides. Both cationic charge as well as hydrophobic segments facilitate interactions with the bacterial cell surface and initiate its disruption. The perturbation in transmembrane potential causes leakage of cytosolic contents followed by cell death. Out of two categories of macromolecules, peptide oligomers and cationic polymers, which have extensively been used as antibacterials, we have elaborated on the current advances made in the area of cationic polymer-based (naturally occurring and commonly employed synthetic polymers and their modified analogs) antibacterial agents. The development of polymer-based antibacterials has helped in addressing challenges posed by the drug-resistant bacterial infections. These polymers provide a new platform to combat such infections in the most efficient manner. This review presents concise discussion on the amphiphilic cationic polymers and their modified analogs having low hemolytic activity and excellent antibacterial activity against array of fungi, bacteria and other microorganisms.


New Mutations and Pathogenesis of Pulmonary Hypertension: Progress and Puzzles in Disease Pathogenesis.
Pulmonary arterial hypertension (PAH) is a complex multifactorial disease with poor prognosis characterized by functional and structural alterations of the pulmonary circulation causing marked increase in pulmonary vascular resistance, ultimately leading to right heart failure and death. Mutations in the gene encoding BMPRII-a receptor for the TGF-β (transforming growth factor-beta) superfamily-account for over 70% of families with PAH and ≈20% of sporadic cases. In recent years, however, less common or rare mutations in other genes have been identified. This review will consider how these newly discovered PAH genes could help to provide a better understanding of the molecular and cellular bases of the maintenance of the pulmonary vascular integrity, as well as their role in the PAH pathogenesis underlying occlusion of arterioles in the lung. We will also discuss how insights into the genetic contributions of these new PAH-related genes may open up new therapeutic targets for this, currently incurable, cardiopulmonary disorder.


The Interplay Between Prenatal Adversity, Offspring Dopaminergic Genes, and Early Parenting on Toddler Attentional Function.
Background: Few studies have explored the complex gene-by-prenatal environment-by-early postnatal environment interactions that underlie the development of attentional competence. Here, we examined if variation in dopamine-related genes interacts with prenatal adversity to influence toddler attentional competence and whether this influence is buffered by early positive maternal behavior. Methods: From the Maternal Adversity, Vulnerability and Neurodevelopment cohort, 134 participants (197 when imputing missing data) had information on prenatal adversity (prenatal stressful life events, prenatal maternal depressive symptoms, and birth weight), five dopamine-related genes (DAT1, DRD4, DRD2, COMT, BDNF), observed maternal parenting behavior at 6 months and parent-rated toddler attentional competence at 18 and 24 months. The Latent Environmental and Genetic Interaction (LEGIT) approach was used to examine genes-by-prenatal environment-by-postnatal environment interactions while controlling for sociodemographic factors and postnatal depression. Results: Our hypothesis of a three-way interaction between prenatal adversity, dopamine-related genes, and early maternal parenting behavior was not confirmed. However, consistent two-way interactions emerged between prenatal adversity and dopamine-related genes; prenatal adversity and maternal parenting behavior, and dopamine-related genes and maternal parenting behavior in relation to toddler attentional competence. Significant interaction effects were driven by the DAT1, COMT, and BDNF genotypes; prenatal stressful life events; maternal sensitivity, tactile stimulation, vocalization, and infant-related activities. Conclusions: Multiple dopamine-related genes affected toddler attentional competence and they did so in interaction with prenatal adversity and the early rearing environment, separately. Effects were already visible in young children. Several aspects of early maternal parenting have been identified as potential targets for intervention.


Identification of phosphorylation sites within the signaling adaptor APPL1 by mass spectrometry.
APPL1 is a membrane-associated adaptor protein implicated in various cellular processes, including apoptosis, proliferation, and survival. Although there is increasing interest in the biological roles as well as the protein and membrane interactions of APPL1, a comprehensive phosphorylation profile has not been generated. In this study, we use mass spectrometry (MS) to identify 13 phosphorylated residues within APPL1. By using multiple proteases (trypsin, chymotrypsin, and Glu C) and replicate experiments of linear ion trap (LTQ) MS and LTQ-Orbitrap-MS, a combined sequence coverage of 99.6% is achieved. Four of the identified sites are located in important functional domains, suggesting a potential role in regulating APPL1. One of these sites is within the BAR domain, two cluster near the edge of the PH domain, and one is located within the PTB domain. These phosphorylation sites may control APPL1 function by regulating the ability of APPL1 domains to interact with other proteins and membranes.


Regulation of prothoracic gland ecdysteroidogenic activity leading to pupal metamorphosis.
The prothoracic glands of early last (fifth) instar larvae of the silkworm are inactive with regard to ecdysteroidogenesis and unresponsive to prothoracicotropic hormone (PTTH) [J. Insect Physiol. 31 (1985) 455]. In an attempt to elucidate the hormonal mechanisms that cause the inactivity, we compared the effects of PTTH, dibutyryl cyclic AMP (dbcAMP), a cAMP phosphodiesterase inhibitor (IBMX), juvenile hormone analogue (JHA) and 20-hydroxyecdysone (20E) on secretory activity of the third, fourth and fifth instar glands. Among the factors examined, feedback inhibition by 20E was indicated to be the most likely factor. Inhibition was moderate in the third and early fourth instars while 20E strongly inhibited the glands of middle fourth instar larvae. The inhibitory effect of 20E was reduced by removal of the brain and corpora allata. Once the glands were suppressed by 20E to the degree of exhibiting neither secretory activity nor responsiveness to PTTH, dbcAMP or IBMX did not elicit ecdysone secretion at all. Thus the feedback inhibition may shut down ecdysteroidogenesis although it is obscure whether it affects the intracellular transductory cascade from the PTTH receptor through cAMP. Taken together, this evidence suggests that inactivity of the gland in the early fifth instar is brought about by feedback inhibition of the glands by 20E occurring in the late fourth instar, and that this inactivity is maintained by the juvenile hormone found in the early fifth instar.


Effects of total fasting in obese women. IV. Response of serum triiodothyronine (T3) and reverse triiodothyronine (rT3) to administered T3.
Serum levels of T4, T3, rT3, RT3U and TSH were estimated in 12 obese women in the course of a 14-day fasting. Seven of these patients were treated with T3 in a daily dose of 60--80 micrograms. Fasting led to a small increase of serum T4, while fasting combined with T3 administration was accompanied by a small decrease of serum T4. Serum T3 decreased in the course of the first 2 days of fasting to 40--50% of initial values and remained at this low level up to the 15th day of fasting. In the T3 treated group a prompt increase of serum T3 was recorded (+80%), followed by a steady decrease, reaching the control values on day 15, in spite of a continuous T3 administration. Serum rT3 in the untreated fasting group steadily increased up to the 12th day (+43%) and then dropped below the control value (-28%). In the treated group after a non-significant increase (+22%), a decrease of serum rT3 was also observed (-42%). Fasting was accompanied by a tendency to increased serum RT3U values, in the T3 treated group no change occurred. TSH in the untreated fasting women remained practically unchanged while T3 administration was accompanied by a tendency to a transient decrease. The heart rate showed a non-significant tendency to increase in the T3 treated group and the last week of fasting was also accompanied in several patients by a larger weight loss. These results suggest that in addition to an enhanced synthesis of rT3 to the detriment of a decreased production of T3 from T4, an increased catabolism of administered T3 and of endogenous rT3 is involved in a strict calorie restriction. An abortive T3-TRH-TSH feed-back may also operate in these conditions.


Stress activation of glutamate neurotransmission in the prefrontal cortex: implications for dopamine-associated psychiatric disorders.
In most psychiatric disorders, stress is the major nongenomic factor that contributes to the expression or exacerbation of acute symptoms, recurrence or relapse after a period of remission, and treatment outcome. Delineation of mechanisms by which stress contributes to these processes is fundamental to understanding the disease process and for improving outcome. In this article, evidence is reviewed to indicate that many central aspects of stress response, including activation of the hypothalmic-pituitary-adrenal (HPA) axis and dopamine neurotransmission, are modulated, and in some cases mediated, by glutamate neurotransmission in the prefrontal cortex (PFC). It is suggested that activation of glutamatergic neurotransmission in the PFC presents a common mechanism by which stress influences normal and abnormal processes that sustain affect and cognition. Although monoamines, in particular dopamine, have been considered the major culprits in the adverse effects of stress in disorders such as addiction and schizophrenia, it is likely that in a vulnerable brain with an underlying PFC pathophysiology, abnormal stress-activated monoaminergic neurotransmission is secondary to anomalies in cortical glutamate neurotransmission. Thus, understanding the contribution of glutamate-mediated processes to stress response through the use of experimental models that involve disrupted PFC function can provide insights to the fundamental pathophysiology of stress-sensitive psychiatric disorders and lead to novel strategies for treatment and prevention.


Isoproterenol-induced current changes in glands of frog skin.
In this study we report on isoproterenol-induced changes in short-circuit current (SCC) that are located in the glands of the frog skin. Following complete inhibition of the SCC with amiloride (10(-4) M), the addition of isoproterenol (9.0 X 10(-7) M) to the serosal side of the skin elicits a significant increase in SCC from 0.6 +/- 0.1 to 3.9 +/- 0.5 microA/cm2 (p less than 0.001). The conductance is doubled, and both effects are blocked completely with the beta-blocking agent propranolol. In the split-skin preparation, in which no glands are present, there is no current response to isoproterenol following amiloride. This indicates that the amiloride-insensitive isoproterenol-stimulated SCC resides in the glands. In the intact skin, removal of sodium (choline replacement) from the serosal (but not the mucosal) medium abolishes this current change. The postisoproterenol current change in amiloride-inhibited skins is also dependent on the presence of chloride in the serosal bath. Preincubation of the amiloride-inhibited skin with ouabain (10(-4) M) for 10 min prior to the addition of isoproterenol reduces the isoproterenol response by 50%. These results indicate that there is a sodium- and chloride-dependent ouabain-sensitive secretory mechanism in the skin glands of the frog that is responsible for the development of the isoproterenol-stimulated SCC observed in the presence of amiloride.


An Evaluation of an Online Brief Mindfulness-Based Intervention in Higher Education: A Pilot Conducted at an Australian University and a British University.
Mental ill health among higher education students is a well-established problem; therefore, it is imperative to implement preventative approaches to support wellbeing. Blended and fully online education programmes widens access for mature or returning students; however, the psychological wellbeing of this sub-group of students is under-researched. Finally, evaluating wellbeing interventions that meet the needs of university students as well as accessible for online students is required. The aim of this study was to evaluate a brief, online and mindfulness-based intervention to assist the self-management of wellbeing and stress for both online and on-campus higher education students. The total sample included 427 participants (96% psychology students) at Monash University, Australia (n=283) and King's College London (n=144), with 152 participants completing the whole study. Participants were allocated to a brief, self-guided, online and mindfulness-based intervention (over the course of one study period; n=297), or to a wait-list control group (n=148). Baseline and end of semester questionnaires included the 14-item Warwick-Edinburgh Mental Wellbeing Scale, 10-item Perceived Stress Scale and the 15-item Mindful Attention Awareness Scale. Regression modelling revealed the intervention condition accounted for up to 12% of the variability in change in student wellbeing, stress and mindfulness between the start and end of semester (when controlling for baseline). These findings support the implementation of a brief, online and asynchronous mindfulness-based intervention for supporting student mental health and psychological wellbeing. An on-going challenge in practice includes engaging and maintaining student engagement in wellbeing initiatives.


The effect of local speaking rate on the perception of quantity in Estonian.
The Estonian language with its elaborate system of contrasts in quantity, whose essentials are described in the paper, is used to investigate human perception of distinctive contrasts in the duration of vowels, consonants and larger units. In the experiments reported, the speaking rate of a preceding or following syllable was manipulated in addition to that of a target V, C or VC sequence that carried a quantity distinction in disyllabic words. The results confirmed that the second syllable in such words, in particular the duration of its vowel, serves as a reference, but they showed segments of additional syllables to contribute in the same direction. The results provided no support for ascribing quantity to any larger units than phonetic segments. Speech rate effects of similar magnitude have been observed in Japanese, while effects of the same kind were found to be smaller in Dutch. These differences may be linked with the functions durational contrasts have in the different languages. It appears that listeners have to adapt more fully to variations in the local speaking rate when there are no additional cues and the functional load of quantity distinctions is high.


Liver biopsy may facilitate pancreatic graft evaluation: Positive association between liver steatosis and pancreatic graft adipose infiltration.
The number of pancreatic transplants has decreased in recent years. Pancreatic grafts have been underutilized compared to other solid grafts. One cause of discard is the macroscopic appearance of the pancreas, especially the presence of fatty infiltration. The current research is aimed at understanding any graft-related association between fatty tissue infiltration of the pancreas and liver steatosis. From August 2013 to August 2014, a prospective cross-sectional clinical study using data from 54 multiple deceased donor organs was performed. Micro- and macroscopic liver steatosis were significantly correlated with the donor body mass index ([BMI]; p=0.029 and p=0.006, respectively). Positive gamma associations between pancreatic and liver macroscopic and microscopic findings (0.98; confidence interval [CI]: 0.95-1 and 0.52; CI 0.04-1, respectively) were observed. Furthermore, comparisons of liver microscopy findings showed significant differences between severe versus absent (p<0.001), severe versus mild (p<0.001), and severe versus moderate classifications (p<0.001). The area under the receiver operating curve was 0.94 for the diagnosis of steatosis by BMI evaluation using a cut-off BMI of 27.5 kg/m2, which yielded 100% sensitivity, 87% specificity, and 100% negative predictive value. We observed a positive association of macroscopic and microscopic histopathological findings in steatotic livers with adipose infiltration of pancreatic grafts.


Specific expression and function of the A-type cytochrome c oxidase under starvation conditions in Pseudomonas aeruginosa.
Pseudomonas aeruginosa has one A-type (caa3) and multiple C-type (cbb3) cytochrome c oxidases as well as two quinol oxidases for aerobic respiration. The caa3 oxidase is highly efficient in creating a proton gradient across the cell membrane, but it is not expressed under normal growth conditions and its physiological role has not been investigated. In the present study, a mutant strain deficient in the coxBA-PA0107-coxC genes encoding caa3 exhibited normal growth under any test conditions, but it had low relative fitness under carbon starvation conditions, indicating that the expression of caa3 is advantageous under starvation conditions. A mutant that lacked four terminal oxidase gene clusters except for the cox genes was unable to grow aerobically because of low expression level of caa3. However, suppressor mutants that grew aerobically using caa3 as the only terminal oxidase emerged after aerobic subculturing. Analyses of the suppressor mutants revealed that a mutation of roxS encoding a sensor kinase of a two-component regulator RoxSR was necessary for the aerobic growth in synthetic medium. Two additional mutations in the 5'-flanking region of coxB were necessary for the aerobic growth in LB medium. Although the expression level of caa3 was higher in the suppressor mutants, their growth rates were lower than when the other terminal oxidases were utilized, suggesting that caa3 was not suited for utilization as the only terminal oxidase. Overexpression of the cox genes also inhibited the aerobic growth of the wild-type strain. These results indicate that caa3 is tightly regulated to be expressed only under starvation conditions at low level and it functions in cooperation with other terminal oxidases to facilitate survival in nutrient starvation conditions.


Fenoprofen in patients with rheumatoid arthritis receiving maintenance gold therapy.
Thirty patients with active definite or classic rheumatoid arthritis (RA), participated in a double-blind crossover study comparing fenoprofen calcium with placebo. All patients were receiving maintenance gold therapy. Fenoprofen was administered in doses ranging from 200 to 800 mg every six hours. The median daily dose was 1.8 gm. During this 16-week trial, most parameters of efficacy showed fenoprofen to be significantly more effective than placebo. Adverse reactions, which were minimal, suggest that fenoprofen will be useful in the management of patients with RA, particularly in those with gastrointestinal intolerance to the usually employed drugs. The longterm effectiveness in RA, and the side effects associated with chronic administration remain to be assessed.


Bioavailability of a novel midazolam gel after intranasal administration in dogs.
To compare the pharmacokinetics of a novel bioadhesive gel formulation of midazolam after intranasal (IN) administration with that of midazolam solution after IN, IV, and rectal administration to dogs. 10 (5 males and 5 females) healthy adult Beagles. Dogs were assigned to 4 treatment groups for a crossover study design. Initially, midazolam solution (5 mg/mL) was administered (0.2 mg/kg) IV to group 1, rectally to group 2, and IN to group 3; a 0.4% hydroxypropyl methylcellulose midazolam gel formulation (50 mg/mL) was administered (0.2 mg/kg, IN) to group 4. Each dog received all 4 treatments; there was a 7-day washout period between subsequent treatments. Blood samples were collected before and after midazolam administration. Plasma concentration of midazolam was determined by use of high-performance liquid chromatography. The peak plasma concentration after IN administration of the gel formulation was significantly higher than that after IN and rectal administration of the solution. Mean ± SD time to peak concentration was 11.70 ± 2.63 minutes (gel IN), 17.50 ± 2.64 minutes (solution IN), and 39 ± 14.49 minutes (solution rectally). Mean bioavailability of midazolam was 70.4% (gel IN), 52.0% (solution IN), and 49.0% (solution rectally). Bioavailability after IN administration of the gel formulation was significantly higher than that after IN and rectal administration of the solution. IN administration of midazolam gel was superior to both IN and rectal administration of midazolam solution with respect to peak plasma concentration and bioavailability.


Superinfection in ducks persistently infected with duck plague virus.
Superinfections with homologous or heterologous strains of duck plague virus resulted in the deaths of birds persistently infected with duck plague virus. Not all birds that were superinfected died. Protection against mortality depended on the route of exposure, strain of the initial duck plague virus, and strain of the superinfecting virus.


Minimal barriers to invasion during human colorectal tumor growth.
Intra-tumoral heterogeneity (ITH) could represent clonal evolution where subclones with greater fitness confer more malignant phenotypes and invasion constitutes an evolutionary bottleneck. Alternatively, ITH could represent branching evolution with invasion of multiple subclones. The two models respectively predict a hierarchy of subclones arranged by phenotype, or multiple subclones with shared phenotypes. We delineate these modes of invasion by merging ancestral, topographic, and phenotypic information from 12 human colorectal tumors (11 carcinomas, 1 adenoma) obtained through saturation microdissection of 325 small tumor regions. The majority of subclones (29/46, 60%) share superficial and invasive phenotypes. Of 11 carcinomas, 9 show evidence of multiclonal invasion, and invasive and metastatic subclones arise early along the ancestral trees. Early multiclonal invasion in the majority of these tumors indicates the expansion of co-evolving subclones with similar malignant potential in absence of late bottlenecks and suggests that barriers to invasion are minimal during colorectal cancer growth.


A parametric fitting algorithm for segmentation of cell images.
This paper presents a parametric fitting algorithm for segmentation of cervical and breast cell images from cytology smears. A parametric elliptical model for cells is introduced and the parameters adjusted to fit the cell shapes while minimizing a cost function. Segmentation results of noisy human cervical cell and textured breast cell images demonstrate that the proposed parametric fitting algorithm is very successful in segmentation of images of both nonoverlapped and overlapped elliptically shaped cells.


Red cell distribution width and hypertensive response to exercise in patients with type 2 diabetes mellitus.
There is no study about hypertensive response to exercise (HRE), which is a marker of unborn hypertension (HT), and red cell distribution width (RDW) association, in diabetic normotensive patients. So, we aimed to investigate any correlation among RDW and HRE in normotensive type 2 diabetic patients. Consecutive type 2 diabetic patients without history of HT and with normal blood pressure (BP) on ambulatory BP monitoring were included to the study. We divided the patients into two groups depending on their peak systolic BP on exercise; HRE (Group 1) or normal response to exercise (Group 2). Data of 75 diabetic patients (51.9 ± 9.7) were analyzed (31 male (48%)). Their mean RDW was 13.11 ± 0.46. Patients with HRE were significantly older than patients without HRE. Smoking was more frequent in Group 2. Gender distribution and body mass index were similar between the groups. Else hemoglobin, hematocrit, red blood cell count and RDW values were not significantly different. Office systolic BP and diastolic BP, daytime and 24-h systolic BP were significantly higher in Group 1 but heart rate was similar between the groups. This study revealed that RDW do not differ between diabetic normotensive patients with HRE or not.


Portal vein encasement predicts neoadjuvant therapy response in liver transplantation for perihilar cholangiocarcinoma protocol.
Survival and recurrence of cancer after liver transplant (LT) for perihilar cholangiocarcinoma (CCA) following neoadjuvant chemoradiotherapy are strongly correlated with the presence of residual CCA in the liver explant. To determine factors predicting response to neoadjuvant therapy using the presence of residual CCA on explant as a surrogate marker. Characteristics of 109 patients having undergone LT for cholangiocarcinoma were abstracted, with attention to parameters hypothesized to influence radiation therapy efficacy. In the multivariable model, the presence of portal vein encasement (OR 11.8; 95% CI: 2.43-57.21; P = 0.002) and MELD score (OR 1.13; 95% CI: 1.02-1.26; P = 0.017) were predictive of residual macroscopic disease (c-statistics 0.78). Oral capecitabine in addition to standard 5-fluorouracil chemotherapy (OR 0.32, 95% CI: 0.14, 0.71; P = 0.006) was independently protective against residual cancer, independent of MELD score. Portal vein encasement was strongly predictive of residual macroscopic disease. Radial tumor diameter did not have greater predictive value than longitudinal diameter, confirming the appropriateness of current protocol selection criteria. No particular tumor morphology predicted better response. Maintenance oral capecitabine following 5-fluorouracil infusion was independently protective against residual disease. Portal vein encasement as a negative prognostic finding should be taken into account to optimize patient selection and management.


Extended exenatide administration enhances lipid metabolism and exacerbates pancreatic injury in mice on a high fat, high carbohydrate diet.
This study expanded upon a previous study in mice reporting a link between exenatide treatment and exocrine pancreatic injury by demonstrating temporal and dose responses and providing an initial mechanistic hypothesis. The design of the present study included varying lengths of exenatide exposure (3, 6 weeks to 12 weeks) at multiple concentrations (3, 10, or 30 µg/kg) with multiple endpoints (histopathology evaluations, immunoassay for cytokines, immunostaining of the pancreas, serum chemistries and measurement of trypsin, amylase, and, lipase, and gene expression profiles). Time- and dose-dependent exocrine pancreatic injury was observed in mice on a high fat diet treated with exenatide. The morphological changes identified in the pancreas involved acinar cell injury and death (autophagy, apoptosis, necrosis, and atrophy), cell adaptations (hypertrophy and hyperplasia), and cell survival (proliferation/regeneration) accompanied by varying degrees of inflammatory response leading to secondary injury in pancreatic blood vessels, ducts, and adipose tissues. Gene expression profiles indicated increased signaling for cell survival and altered lipid metabolism in exenatide treated mice. Immunohistochemistry supported gene expression findings that exenatide caused and/or exacerbated pancreatic injury in a high fat diet environment potentially by further increasing high fat diet exacerbated lipid metabolism and resulting oxidative stress. Further investigation is required to confirm these findings and determine their relevance to human disease.


An intracellular structure in Bacteroides fragilis.
An unusual intracellular circular or oval structure was discovered during a morphological study of Bacteroides fragilis by electronmicroscopy. The structure is complex, composed of multiple concentric layers surrounding a central region. The dimensions and profile are consistent in all the sections observed. The sections illustrated may represent various stages in the morphogenesis of the complete structure.


Factors associated with diffusely increased renal uptake of Tc-99m diphosphono-propanedicarboxylic acid on bone scintigraphy in patients with end-stage renal disease.
We aimed to determine the factors contributing toward diffusely increased renal uptake on bone scintigraphy using technetium-99m (Tc-99m) diphosphono-propanedicarboxylic acid (DPD) in patients with end-stage renal disease. One-hundred and forty-three bone scintigraphies, performed between June 2007 and July 2013, in 135 patients with chronic kidney disease were analyzed retrospectively, including 22 bone scintigraphies (15 patients; eight women; seven men) with glomerular filtration rates less than 15 ml/min/1.73 m. Patients with nephrocalcinosis were excluded. The clinical records of medication and underlying disease were reviewed retrospectively. The presence of renal artery calcification was visually estimated and the Hounsfield unit (HU) of renal parenchyma was measured by abdominal and pelvic nonenhanced computed tomography. Two patients underwent peritoneal dialysis and 13 underwent hemodynamic dialysis. Diffusely increased renal Tc-99m DPD uptake was observed on 15 of 22 (68%) bone scintigraphy images. Laboratory test results were not significantly different between the patients with or without increased renal uptake. A history of inflammatory conditions or antibiotic administration did not correlate with kidney visualization. The significant factor for diffuse renal tracer uptake on multivariate analysis was renal artery calcification (odds ratio: 18.42; 95% confidence interval: 2.01-79.43; P<0.001). The mean HU values were significantly higher in patients with diffuse renal tracer uptake (P=0.04) and renal artery calcification (P=0.02). Small renal arteriolar calcification and a higher HU value of renal parenchyma could be associated with diffusely increased renal Tc-99m DPD uptake in patients with end-stage renal disease.


Truncation of the otoferlin transmembrane domain alters the development of hair cells and reduces membrane docking.
Release of neurotransmitter from sensory hair cells is regulated by otoferlin. Despite the importance of otoferlin in the auditory and vestibular pathways, the functional contributions of the domains of the protein have not been fully characterized. Using a zebrafish model, we investigated a mutant otoferlin with a stop codon at the start of the transmembrane domain. We found that both the phenotype severity and the expression level of mutant otoferlin changed with the age of the zebrafish. At the early developmental time point of 72 h post fertilization, low expression of the otoferlin mutant coincided with synaptic ribbon deficiencies, reduced endocytosis, and abnormal transcription of several hair cell genes. As development proceeded, expression of the mutant otoferlin increased, and both synaptic ribbons and hair cell transcript levels resembled wild type. However, hair cell endocytosis deficits and abnormalities in the expression of GABA receptors persisted even after up-regulation of mutant otoferlin. Analysis of membrane-reconstituted otoferlin measurements suggests a function for the transmembrane domain in liposome docking. We conclude that deletion of the transmembrane domain reduces membrane docking, attenuates endocytosis, and results in developmental delay of the hair cell.


How different pruning severities alter shoot structure: a modelling approach in young 'Nonpareil' almond trees.
Axillary meristem fate patterns along shoots, also referred to as shoot structure, appear to be fairly consistent among trees within a genotype growing under similar conditions. Less is known about shoot structural plasticity following external manipulations, such as pruning. The aim of this study on almond (Prunus dulcis (Mill.)) shoots was to investigate how pruning severity affects the structure of 1-year-old shoots that grew after pruning (regrowth shoots), the 2-year-old portion of shoots that remained from the previous year's growth after pruning (pruned shoots), and whether regrowth shoots reiterate the structure of the original 1-year-old shoots before pruning. Three pruning severities were imposed and the structures along the different shoots were assessed by building hidden semi-Markov models of axillary meristem fates. The structures of regrowth and pruned shoots depended on pruning severity, but maintained some of the original shoot characteristics. Regrowth shoots developed more complex structures with severe pruning, but had simpler structures than original shoots indicating progressive simplification with tree age. Pruned shoot structures were affected by the severity of pruning, by the structure when the shoots were 1 year old, and probably by local competition among buds. Changes in structure due to pruning can be modelled and be predictable.


Social functioning difficulties in ADHD: association with PDD risk.
Although social difficulties are a common feature of Attention-Deficit Hyperactivity Disorder (ADHD), little is known about the diversity of social problems, their etiology, or their relationship to disorders of social behavior, such as autism or Pervasive Developmental Disorder (PDD). In 379 children and adolescents with ADHD, social functioning was assessed using the Child Behavior Checklist (Achenbach, 1991). Factor analysis and structural equation modeling revealed two factors that we labeled Peer Rejection and Social Immaturity. A factor reflecting ;PDD risk' was defined from eight items of a separate screening instrument for PDD and examined for its association with these two social factors. There was a significant association with both factors, but the association was much stronger for the Social Immaturity (Standardized Beta [beta ] = .51) than Peer Rejection (beta = .29) factors. Social Immaturity was also associated with a greater number of hyperactive symptoms while high Peer Rejection was associated with increased aggression and lower IQ in the ADHD children.


Inherently magnetic hydrogel for data storage based on the magneto-optical Kerr effect.
In this data explosion age, a large amount of data is generated every day. Such a fast data growth has aroused great interest in the field of data storage. Conventional data storage materials are mainly composed of hard and brittle materials but they may break in the case of mechanical operations, causing irreversible data loss. In this work, efforts have been devoted to fabricating a flexible and stretchable double network hydrogel for data storage based on the magneto-optical Kerr effect. The hydrogel possesses a storage modulus of over 104 Pa and remains unbroken under a strain of 3000%. The surface of the hydrogel is patterned with diamagnetic parts and paramagnetic parts alternately. When placed under a magnetic field, the surface of the hydrogel reflects the incident laser beam and changes the polarization plane of the reflected light. The outstanding flexibility and inherent magnetic properties of this hydrogel lay the groundwork for data storage and guarantee data safety.


Repeated sauna therapy reduces urinary 8-epi-prostaglandin F(2alpha).
We have reported that repeated sauna therapy improves impaired vascular endothelial function in a patient with coronary risk factors. We hypothesized that sauna therapy decreases urinary 8-epi-prostaglandin F(2alpha) (PGF(2alpha)) levels as a marker of oxidative stress and conducted a randomized, controlled study. Twenty-eight patients with at least one coronary risk factor were divided into a sauna group (n = 14) and non-sauna group (n = 14). Sauna therapy was performed with a 60 degrees C far infrared-ray dry sauna for 15 minutes and then bed rest with a blanket for 30 minutes once a day for two weeks. Systolic blood pressure and increased urinary 8-epi-PGF(2alpha) levels in the sauna group were significantly lower than those in the non-sauna group at two weeks after admission (110 +/- 15 mmHg vs 122 +/- 13 mmHg, P < 0.05, 230 +/- 67 pg/mg x creatinine vs 380 +/- 101 pg/mg x creatinine, P < 0.0001, respectively). These results suggest that repeated sauna therapy may protect against oxidative stress, which leads to the prevention of atherosclerosis.


The Roles of Time and Change in Situations.
Rauthmann, Sherman and Funder have made a landmark contribution to situation research in the target article of this issue. However, we propose that their work overlooks the need to incorporate a developmental perspective. This includes the separate but related issues of time and change. Situations often unfold over long periods of time, can bleed together, and are not time-delimited in the way traditional laboratory experiments define them. Moreover, individuals systematically change over time (lifespan development) and their reactions to situations, as well as their personality-situation transactions, develop in tandem.


[Fecal microbiota transplantation, a novel therapy for recurrent Clostridium difficile infection].
Clostridium difficile infection is caused by a disturbance of the gut microbiota, often resulting from the use of antibiotics. Among a sub group of patients with this disorder, treatment with antibiotics is not effective. They develop a chronic, recurrent infection. Such patients can be treated with a fecal microbiota transplantation (FMT), or fecal transplantation. The crucial steps for safe application of fecal transplantation are central donor selection and screening. To optimise safety and to guarantee the availability of donor feces for fecal transplantation, the Nederlandse Donor Feces Bank (Dutch Donor Feces Bank) was established. At this facility, ready-to-use, screened donor feces can be ordered for patients with (recurrent) Clostridium difficile infections, who can then be treated at their own hospital.


Exploring morphometric frontiers: A comprehensive study of otolith growth patterns in brown comber Serranus hepatus (Linnaeus, 1758).
Otoliths are widely employed in marine sciences to gain insights into fish growth, age, migrations, and population structure. This study investigates the relationships between morphometric measurements, otolith characteristics, and length size patterns in the brown comber (Serranus hepatus) from the Gulf of Cádiz, a species discarded in artisanal trawl fisheries. Our findings reveal significant changes in otolith shape indices as fish grow, with symmetry observed between left and right otolith measurements. Otolith size is found to be related to fish size, supporting its use in estimating body length at different life stages. Otolith shape analysis has potential applications in stock identification, detecting catch misreporting, and studying marine predator diets. Combining otolith shape analysis with other data types can clarify relationships among taxa and inform spatial management strategies, contributing to the long-term sustainability of fish populations and the assessment of the impact of management strategies on fish size and growth. This study enhances our understanding of the broader implications of morphometric and otolith analyses in fisheries research and supports the development of more sustainable fisheries management practices.


CTRP3 inhibits myocardial fibrosis through the P2X7R-NLRP3 inflammasome pathway in SHR rats.
Reducing hypertensive myocardial fibrosis is the fundamental approach to preventing hypertensive ventricular remodelling. C1q/TNF-related protein-3 (CTRP3) is closely associated with hypertension. However, the role and mechanism of CTRP3 in hypertensive myocardial fibrosis are unclear. In this study, we aimed to explore the effect of CTRP3 on hypertensive myocardial fibrosis and the potential mechanism. WKY and SHR rats were employed, blood pressure, body weight, heart weight, H/BW were measured, and fibrotic-related proteins, CTRP3 and Collagen I were tested in myocardium at 12 and 20 weeks by immunohistochemical staining and Western blotting, respectively. The results showed that compared with the WKY, SBP, DBP, mean arterial pressure and heart rate (HR) were all significantly increased in SHR at 12 and 20 weeks, while heart weight and H/BW were only increased at 20 weeks. Meanwhile, CTRP3 decreased, while Collagen I increased significantly in the SHR rat myocardium at 20 weeks, which compared to the WKY. Moreover, the expression of α-SMA increased from 12 weeks, Collagen I/III and MMP2/9 increased and TIMP-2 decreased until 20 weeks. In order to explore the function and mechanism of CTRP3 in hypertensive fibrosis, Angiotensin II (Ang II) was used to induce hypertension in primary neonatal rat cardiac fibroblasts in vitro. CTRP3 significantly inhibited the Ang II induced activation of fibrotic proteins, purinergic 2X7 receptor (P2X7R)-NLRP3 inflammasome pathway. The P2X7R agonist BzATP significantly exacerbated Ang II-induced NLRP3 inflammasome activation, which was decreased by the P2X7R antagonists A43079, CTRP3 and MCC950. CTRP3 expression was decreased in the myocardium of SHR rats, and exogenous CTRP3 inhibited Ang II-induced fibrosis in cardiac fibroblasts by regulating the P2X7R-NLRP3 inflammasome pathway, suggesting that CTRP3 is a potential drug for alleviating myocardial fibrosis in hypertensive conditions.