Trends in the Distribution of Opioids in Puerto Rico, 1999-2013.
Limited information has been published about opioid prescribing practices in Puerto Rico. The objective of this study was to create baseline trends of opioids distributed over a period of fourteen years in Puerto Rico. We examined data from the U.S. Drug Enforcement Administration's Automation of Reports and Consolidated Orders System (ARCOS) for the period 1999-2013. ARCOS data reflects the amount of controlled substances legally dispensed. Analyses include the distribution of opioids (in morphine milligram equivalent kg per 10,000 persons) by year and entity (pharmacy, hospital, practitioner). The distribution of four drugs (fentanyl, hydromorphone, methadone, oxycodone) increased over 100% between 1999 and 2013. The distribution of two drugs (hydrocodone and meperidine) declined between 1999 and 2013. Oxycodone distribution grew from 0.13 MME kg grams per 10,000 persons in 1999 to 0.29 MME kg in 2013. ARCOS data showed that the overall amount of opioid pain relievers distributed in Puerto Rico increased by 68% between 1999 and 2013. Currently, prescription opioid pain reliever overdose deaths in Puerto Rico do not appear to be skyrocketing as they are in the mainland U.S. However, the ongoing problem with prescription opioid pain reliever overdoses in certain areas should serve as a warning to monitor consumption of opioid pain relievers, as well as changes in prescription drug abuse, overdoses, and deaths.


Loop-mediated isothermal amplification of DNA (LAMP): a new diagnostic tool lights the world of diagnosis of animal and human pathogens: a review.
Diagnosis is an important part in case of animal husbandry as treatment of a disease depends on it. Advancement in molecular biology has generated various sophisticated tools like Polymerase Chain Reaction (PCR), its versions along with pen-side diagnostic techniques. Every diagnostic test however has both advantages and disadvantages; PCR is not an exception to this statement. To ease the odds faced by PCR several non-PCR techniques which can amplify DNA at a constant temperature has become the need of hour, thus generating a variety of isothermal amplification techniques including Nucleic Acid Sequence-Based Amplification (NASBA) along with Self-Sustained Sequence Replication (3SR) and Strand Displacement Amplification (SDA) and Loop mediated isothermal amplification (LAMP) test. LAMP stands out to be a good and effective diagnostic test for empowering in developing countries as it does not require sophisticated equipments and skilled personnel and proves to be cost-effective. Performance of LAMP mainly relies on crafting of six primers (including 2 loop primers) ultimately accelerating the reaction. LAMP amplifies DNA in the process pyrophosphates are formed causing turbidity that facilitates visualisation in a more effective way than PCR. The Bst and Bsm polymerase are the required enzymes for LAMP that does not possess 5'-3' exonuclease activity. Results can be visualized by adding DNA binding dye, SYBR green. LAMP is more stable than PCR and real-time PCR. Non-involvement of template DNA preparation and ability to generate 10(9) copies of DNA are added benefits that make it more effective than NASBA or 3SR and SDA. Thus, it fetches researcher's interest in developing various versions of LAMP viz., its combination with lateral flow assay or micro LAMP and more recently lyophilized and electric (e) LAMP. Availability of ready to use LAMP kits has helped diagnosis of almost all pathogens. LAMP associated technologies however needs to be developed as a part of LAMP platform rather than developing them as separate entities. This review deals with all these salient features of this newly developed tool that has enlightened the world of diagnosis.


Hippocampal adequacy versus functional reserve: predicting memory functions following temporal lobectomy.
While decrements in memory following temporal lobectomy for intractable epilepsy remain a major neurocognitive complication of surgical intervention, it has been difficult to identify patients who are most at risk for developing decrements in their memory functions following surgery. Two different models of hippocampal functioning have been posited to account for postsurgical memory losses: hippocampal reserve versus functional adequacy. This article briefly examines the basis of the traditional model of hippocampal reserve, which suggests that it is the reserve or capacity of the contralateral hippocampus to support memory after surgery that determines whether changes in memory function will be observed. A more in-depth review of emerging data from studies of function (baseline neuropsychological testing and intracarotid amytal procedure) and structure (histologic cell densities and MRI volumetrics) supporting a functional adequacy model of hippocampal functioning is then presented. Data from these latter studies provide strong convergent support for the view that the risk for memory decrements following mesial temporal lobe epilepsy surgery is inversely related to the functional adequacy of the tissue to be resected. Data reviewed here have broad implications for the field of neuropsychology. Not only do the data reported in this article challenge some of the basic tenets of our understanding of the role of the hippocampus in memory functioning, but they illustrate the importance of ongoing research, critical reevaluation of basic concepts, and the importance of self-scrutiny in light of new technologies.


Tobramycin in bronchial secretions.
The penetration of tobramycin sulfate (nebramycin factor 6) into normal lower respiratory tract secretions was studied in a dog model. After a 1.7 mg/kg intravenous dose of tobramycin, the antibiotic was detected promptly in bronchial secretions, but the peak bronchial secretion concentration was not usually reached until 2 h after injection (mean level 1.15 mug/ml). The maximum tobramycin concentration in bronchial fluids was equal to the mean inhibitory concentrations that have been reported for about 80% of Pseudomonas isolates.


Low-dose daily aspirin reduces topical minoxidil efficacy in androgenetic alopecia patients.
Topical minoxidil is the only US FDA approved OTC drug for the treatment of androgenetic alopecia (AGA). Minoxidil is a pro-drug converted into its active form, minoxidil sulfate, by the sulfotransferase enzymes in the outer root sheath of hair follicles. Previously, we demonstrated that sulfotransferase activity in hair follicles predicts response to topical minoxidil in the treatment of AGA. In the human liver, sulfotransferase activity is significantly inhibited by salicylic acid. Low-dose OTC aspirin (75-81 mg), a derivative of salicylic acid, is used by millions of people daily for the prevention of coronary heart disease and cancer. It is not known whether oral aspirin inhibits sulfotransferase activity in hair follicles, potentially affecting minoxidil response in AGA patients. In the present study, we determined the follicular sulfotransferase enzymatic activity following 14 days of oral aspirin administration. In our cohort of 24 subjects, 50% were initially predicted to be responders to minoxidil. However, following 14 days of aspirin administration, only 27% of the subjects were predicted to respond to topical minoxidil. To the best of our knowledge, this is the first study to report the effect of low-dose daily aspirin use on the efficacy of topical minoxidil.


Surface stress, thickness, and mass of the first few layers of polyelectrolyte.
The effects of surface stress and mass loading upon the adsorption of polyelectrolytes onto flexible silicon micromechanical cantilever sensors (MCSs) were studied in situ. A self-assembled monolayer of 2-mercaptoethylamine chloride (2-MEA) on gold was used to achieve single-side adsorption on the MCS. Such a preparation gave a positive surface potential, whereas a bare SiOx surface gave a negative surface potential. Wide scan X-ray photoelectron spectroscopy confirmed that the adsorption of polystyrenesulfonate (PSS) and polyallylamine hydrochloride (PAH) followed the general rule expected from the electrostatic interaction between the substrate and the polyelectrolyte, whereas the adsorption polyethyleneimine (PEI) did not. The adsorption of PAH on SiO(x) from a 3 mM water solution containing 1 M NaCl was associated with a deflection of the MCS toward the polyelectrolyte monolayer (tensile surface stress) owing to the hydrogen bonding between neighboring amino groups. Here, a surface stress change of 1.4 +/- 0.1 N/m was estimated. The adsorption of PSS from a 3 mM water solution containing 1 M NaCl on a 2-MEA surface induced a deflection of the MCS away from the polyelectrolyte layer (compressive stress), toward the SiO(x) side. Here, a surface stress change of 3.1 +/- 0.3 N/m was determined. The formation of a PAH layer on top of the PSS layer resulted in a deflection of the MCS toward the PAH layer. This indicated that the adjacent PSS layer was deswelling, corresponding to a surface stress change of 0.5 +/- 0.1 N/m.


Carbon Nanotube Yarn for Fiber-Shaped Electrical Sensors, Actuators, and Energy Storage for Smart Systems.
Smart systems are those that display autonomous or collaborative functionalities, and include the ability to sense multiple inputs, to respond with appropriate operations, and to control a given situation. In certain circumstances, it is also of great interest to retain flexible, stretchable, portable, wearable, and/or implantable attributes in smart electronic systems. Among the promising candidate smart materials, carbon nanotubes (CNTs) exhibit excellent electrical and mechanical properties, and structurally fabricated CNT-based fibers and yarns with coil and twist further introduce flexible and stretchable properties. A number of notable studies have demonstrated various functions of CNT yarns, including sensors, actuators, and energy storage. In particular, CNT yarns can operate as flexible electronic sensors and electrodes to monitor strain, temperature, ionic concentration, and the concentration of target biomolecules. Moreover, a twisted CNT yarn enables strong torsional actuation, and coiled CNT yarns generate large tensile strokes as an artificial muscle. Furthermore, the reversible actuation of CNT yarns can be used as an energy harvester and, when combined with a CNT supercapacitor, has promoted the next-generation of energy storage systems. Here, progressive advances of CNT yarns in electrical sensing, actuation, and energy storage are reported, and the future challenges in smart electronic systems considered.


Impact of ileostomy-related complications on the multidisciplinary treatment of rectal cancer.
Radical resection is the primary treatment for rectal cancer. When anastomosis is possible, a temporary ileostomy is used to decrease morbidity from a poorly healed anastomosis. However, ileostomies are associated with complications, dehydration, and need for a second operation. We sought to evaluate the impact of ileostomy-related complications on the treatment of rectal cancer. We conducted a retrospective study of patients who underwent sphincter-preserving surgery between January 2005 and December 2010 at a tertiary cancer center. The primary outcome was the overall rate of ileostomy-related complications. Secondary outcomes included complications related to ileostomy status, ileostomy closure, anastomotic complications at primary resection, rate of stoma closure, and completion of adjuvant chemotherapy assessed by multivariate logistic regression. Of 294 patients analyzed, 32% (n = 95) were women. Two hundred seventy-one (92%) received neoadjuvant chemoradiation. The median tumor distance from the anal verge was 7 cm (interquartile range 5-10 cm). Two hundred eighty-one (96%) underwent stoma closure at a median of 7 months (interquartile range 5.4-8.3 months). The most common complication related to readmission was dehydration (n = 32-11%). Readmission within 60 days of primary resection was associated with delay in initiating adjuvant chemotherapy (odds ratio 3.01, 95% confidence interval 1.42-6.38, p = 0.004). Diverting ileostomies created during surgical treatment of rectal cancers are associated with morbidity; however, this is balanced against the risk of anastomosis-related morbidity at rectal resection. Given the potential benefit of fecal diversion, patient-oriented interventions to improve ostomy management, particularly during adjuvant chemotherapy, can be expected to yield marked benefits.


Adaptive relaxation labeling.
Current implementation of probabilistic relaxation labeling (PRL) is based on stationary compatibility coefficients (SCC's). Such labeling frequently diverges from an achieved minimum labeling error. In this correspondence it is shown that by having nonstationary compatibility coefficients (NSCC's) the PRL stabilizes about the minimum error which is obtained during the early iterations. Also, a noniterative labeling algorithm which uses NSCC and has a performance similar to that of the modified PRL is suggested.


[Detection of cytomegalovirus by the immunoperoxidase technique and viral isolation].
An Immunoperoxidase assay was applied to detect early antigens of cytomegalovirus (CMV) in 150 urine samples from immunocompromised patients, using the commercial available monoclonal antibody against CMV E13. The detection of early antigen by IP (IPCMV) is compared to the conventional cell culture isolation regarding specificity and sensitivity in order to evaluate is usefulness in the diagnostic of CMV infections. The IPCMV showed a sensitivity of 89.8% and a specificity of 91.3% when compared to the isolation method. The great advantage of the IPCMV is based on the shorter time results are achieved, since 48-72 Hs can be enough to provide evidence of CMV infection, while in the isolation technique cytopathogenic effect was present around 14 days after sample inoculation.


Effect of elevated carbon-dioxide on plant growth, physiology, yield and seed quality of chickpea (Cicer arietinum L.) in Indo-Gangetic plains.
In the present scenario of climate change with constantly increasing CO2 concentration, there is a risk of altered crop performance in terms of growth, yield, grain nutritional value and seed quality. Therefore, an experiment was conducted in open top chamber (OTCs) during 2017-18 and 2018-19 to assess the effect of elevated atmospheric carbondioxide (e[CO2]) (600 ppm) on chickpea (cv. JG 14) crop growth, biomass accumulation, physiological function, seed yield and its quality in terms of germination and vigour. The e[CO2] treatment increased the plant height, leaf and stem biomass over ambient CO2 (a[CO2]) treatment. The e[CO2] increased seed yield by 11-18% which was attributed to an increase in the number of pods (6-10%) and seeds plant-1 (8-9%) over a[CO2]. However, e[CO2] reduced the seed protein (7%), total phenol (13%) and thiobarbituric acid reactive substances (12%) and increased the starch (21%) and water uptake rate as compared to seeds harvested from a[CO2] environment. Exposing chickpea plant to e[CO2] treatment had no impact on germination and vigour of the harvested seeds. Also, the physical attributes, total soluble sugar and antioxidant enzymes activities of harvested seeds were comparable in a[CO2] and e[CO2] treatment. Hence, the experimental findings depict that e[CO2] upto 600 ppm could add to the growth and productivity of chickpea in a sub-tropical climate with an implication on its nutritional quality of the produce.


Immunisation of colorectal cancer patients with autologous tumour cells.
Patients with colorectal cancer were entered into a clinical phase I trial of immunotherapy with an autologous tumour cell/bacillus Calmette-Guerin (BCG) vaccine. We attempted to describe the possible effects and side effects of the immunisation, and further to investigate whether expression of immune-response-related surface molecules on the tumour cells in the vaccine correlated with survival. The first and second vaccine comprised of 107 irradiated tumour cells mixed with BCG, the third of irradiated tumour cells only. Thirty-nine patients were considered, but only 6 patients fulfilled the criteria for inclusion. No serious side effects were observed. With three years of observation time, two patients are healthy, while the rest have had recurrence, and two of them have died. In all vaccines, all tumour cells expressed HLA class I, some expressed HLA class II and none expressed CD80. There was an inverse relation between survival and HLA class II expression. This highlights an essential problem, in the absence of CD80 expression the expression of HLA class II may induce anergy. In future attempts to develop improved vaccines this problem should be addressed.


Using deep RNA sequencing for the structural annotation of the Laccaria bicolor mycorrhizal transcriptome.
Accurate structural annotation is important for prediction of function and required for in vitro approaches to characterize or validate the gene expression products. Despite significant efforts in the field, determination of the gene structure from genomic data alone is a challenging and inaccurate process. The ease of acquisition of transcriptomic sequence provides a direct route to identify expressed sequences and determine the correct gene structure. We developed methods to utilize RNA-seq data to correct errors in the structural annotation and extend the boundaries of current gene models using assembly approaches. The methods were validated with a transcriptomic data set derived from the fungus Laccaria bicolor, which develops a mycorrhizal symbiotic association with the roots of many tree species. Our analysis focused on the subset of 1501 gene models that are differentially expressed in the free living vs. mycorrhizal transcriptome and are expected to be important elements related to carbon metabolism, membrane permeability and transport, and intracellular signaling. Of the set of 1501 gene models, 1439 (96%) successfully generated modified gene models in which all error flags were successfully resolved and the sequences aligned to the genomic sequence. The remaining 4% (62 gene models) either had deviations from transcriptomic data that could not be spanned or generated sequence that did not align to genomic sequence. The outcome of this process is a set of high confidence gene models that can be reliably used for experimental characterization of protein function. 69% of expressed mycorrhizal JGI "best" gene models deviated from the transcript sequence derived by this method. The transcriptomic sequence enabled correction of a majority of the structural inconsistencies and resulted in a set of validated models for 96% of the mycorrhizal genes. The method described here can be applied to improve gene structural annotation in other species, provided that there is a sequenced genome and a set of gene models.


iQuant2: Software for Rapid and Quantitative Imaging Using Laser Ablation-ICP Mass Spectrometry.
We report on the development of a software program named iQuant2 which creates visual images from two-dimensional signal intensity data obtained by a laser ablation-ICP-mass spectrometry (LA-ICPMS) technique. Time-resolved signal intensity profiles can be converted to position resolved signal intensity data based on the rastering rate (μm s-1) of the laser ablation. Background signal intensities obtained without laser ablation (gas blank) are used as the background, and all of the blank-subtracted intensity data can be used for the imaging analysis. With this software, deformation of the created image can be corrected visually on a PC screen. The line profile analysis between the user-selected points can be observed using the iQuant2 software. To accomplish this, data points on the profile line were automatically calculated based on the interpolation between the analysis points. The resulting imaging data can be exported and stored as JPEG, BMP or PNG formats for further processing. Moreover, a semi-quantitative analysis can be made based on the coupling of the external correction of the RSF (relative sensitivity factor) using NIST SRM610 with normalization of the corrected signal intensity data being 100%. The calculated abundance data for major elements are in reasonable agreement with the values obtained by electron probe micro analyzer (EPMA). With the software developed in this study, both the rapid imaging and semi-quantitative determinations can be made.


Which Th pathway is involved during late stage amoebic gill disease?
Amoebic gill disease (AGD) is an emerging disease in North European Atlantic salmon (Salmo salar Linnaeus 1758) aquaculture caused by the amoeba Paramoeba perurans. The host immune response to AGD infection is still not well understood despite past attempts to investigate host-pathogen interactions. With the significant increase in our knowledge of cytokine genes potentially involved in Th responses in recent years, we examined their involvement in this disease using Atlantic salmon post-smolts sampled 3 weeks after exposure to either 500 or 5000 cells/l P. perurans. Gene expression analysis of cytokines potentially involved in the different Th pathways was performed on the first gill arch including the interbranchial lymphoid tissue (ILT). Th1, Th17 and Treg pathways were found to be significantly down regulated, mainly in samples from fish given the higher dose. In contrast, the Th2 pathway was found to be significantly up regulated by both infection doses. Correlation analysis of the gene expression data and the P. perurans load, assessed by real time RT-PCR of the 18S rRNA, was also performed. In humans, Th2 driven responses are characterized by the production of IgE, which in the majority of worm infections results in the generation of a Th2-mediated response and directs the immune system away from a Th1 inflammatory response. The present results seen during late stage AGD suggest that either an immune evasion strategy, similar to the responses driven by helminthic parasites to avoid cell-mediated killing mechanisms, or an allergic reaction caused by the parasite, is occurring.


Day-case tonsillectomy: practical solution or practical impossibility.
The use of day case surgery is on the rise. In order to improve efficiency and reduce cost, it has been proposed that tonsillectomy could be undertaken as a day-case procedure in Ireland. A retrospective, chart-based study was carried out. The medical and social criteria of all patients who underwent tonsillectomy during a twelve-month period were evaluated. Individual, local and national factors were identified and international comparisons were made. Of 161 patients included, 43 (27%) were considered suitable for day case tonsillectomy (DCT). The distance/time criteria from hospital excluded 64% of patients. The diagnosis of obstructive sleep apnoea was the single most common medical reason for exclusion. Support structures were deficient. Local factors must be considered before any policy or targets are developed for DCT. Patient safety is the fundamental tenet. Currently, the infrastructure and the support required for a patient-focused, safe efficient DCT are deficient, and need investment.


Heparin oligosaccharides enhance tissue-type plasminogen activator: a correlation between oligosaccharide length and stimulation of plasminogen activation.
The rate of plasminogen (Pg) activation by tissue-type Pg activator (t-PA) is enhanced by heparin-derived oligosaccharides. Kinetic analysis of the effects of heparin oligosaccharides, ranging in size from di- to dodecasaccharides, on Pg activation demonstrates that stimulation of the reaction is dependent on the size of the heparin oligosaccharides. Di- and tetrasaccharides enhance the activation through 2-fold increases in kcat and 4-fold decreases in Km. Hexasaccharide and larger oligosaccharides stimulate the reaction by increasing the kcat by as much as 4-fold, but do not affect the Km. Previous experiments have shown that lipoprotein(a) [Lp(a)] inhibits Pg activation by t-PA, but only in the presence of a template which enhances t-PA activity such as fibrinogen fragments or intact heparin. Similiarly, Lp(a) inhibits the enhancement of t-PA activity by the larger heparin oligosaccharides but has no effect on t-PA activity in the presence of di- and tetrasaccharides. The results of this study when considered with our previous observations (Edelberg & Pizzo, 1990) suggest that the enhancement in Pg activation by the smaller oligosaccharides is mediated exclusively via binding to t-PA while the larger oligosaccharides may interact with both t-PA and Pg. Furthermore, studies of Pg activation in the presence of both heparin oligosaccharides and fibrinogen fragments demonstrate that t-PA is stimulated preferentially by fibrinogen fragments.


Complex segregation analysis of deafness in Dalmatians.
To use pedigree analysis to evaluate the feasibility of a major locus model for deafness in Dalmatians. 605 purebred Dalmatians from 42 families. Hearing loss was evaluated through the brainstem auditory-evoked response. Dogs were classified into mutually exclusive categories: normal hearing, unilaterally deaf, or bilaterally deaf. Information was collected on sex, coat color, presence or absence of a color patch at birth, and eye color. Statistical analyses were performed by use of regressive logistic models designed for complex segregation analysis. Genetic correlations among eye color, deafness, and color patch were estimated. Prevalence of hearing loss was 11% for dogs classified as unilaterally deaf and 5% for dogs that were bilaterally deaf. Complex segregation analysis detected statistical evidence of a single allele with an expected frequency of 0.21 that had an effect on the prevalence of deafness. Results of analyses suggested that this locus cannot completely explain the inheritance and incidence of deafness in Dalmatians. Genetic correlation estimates among deafness, eye color, and color patch revealed strong interrelationships among these characteristics. To reduce the incidence of hearing loss in Dalmatians, unilaterally deaf, blue-eyed dogs should not be considered as potential parents.


Dose correction strategy for the optimization of volumetric modulated arc therapy.
Dose calculation during optimization of volumetric modulated arc therapy (VMAT) is necessarily simplified to keep computation time manageably low; however the approximations used in the scatter dose calculation lead to discrepancy with more accurate dose calculation following optimization. The purpose of this study is to develop a dose correction strategy in optimization that can minimize the disagreement. VMAT delivery is modeled using a number of static equispaced beams. Dose correction factors (C(ij)) are associated with each beam i and point j inside the region of interest. C(ij) is calculated as the ratio of dose obtained from the full scatter dose calculation over that from the partial scatter dose calculation in optimization. VMAT optimization algorithm is a multiple resolution approach. The dose correction factors are calculated at the beginning of each resolution and applied as multiplicative corrections to the partial scatter dose during optimization. Clinical cases for brain, prostate, paraspinal, and esophagus are utilized to evaluate the method. Treatment plans created with and without the correction scheme are normalized such that the complication rates of organs at risk (OARs) are comparable. The resulting planning target volume (PTV) mean doses are used to compare plan quality. The difference between the dose calculated at the end of optimization and at the end of the final forward dose calculation is reduced from 7% and 5% for the PTV and OAR mean doses without correction to approximately 1% with correction. Applying dose correction during optimization saves planners 2-4 h in average in treatment planning, and has a positive impact on plan quality, evidenced by a noticeably higher PTV mean dose: 2.1%, 2.4%, 0.5%, and 9.3% of the corresponding prescription dose in the brain, esophagus, prostate, and paraspinal cases, respectively. When dose correction is applied during optimization, dose discrepancies between optimization and full dose calculation are reduced. Integrating dose correction in VMAT optimization allows planners to adjust the optimization constraints more easily and confidently during optimization and has the potential to improve plan quality.


The freeze-all strategy versus agonist triggering with low-dose hCG for luteal phase support in IVF/ICSI for high responders: a randomized controlled trial.
Does the freeze-all strategy in high-responders increase pregnancy rates and improve safety outcomes when compared with GnRH agonist triggering followed by low-dose hCG intensified luteal support with a fresh embryo transfer? Pregnancy rates after either fresh embryo transfer with intensified luteal phase support using low-dose hCG or the freeze-all strategy did not vary significantly; however, moderate-to-severe ovarian hyperstimulation syndrome (OHSS) occurred more frequently in the women who attempted a fresh embryo transfer. Two strategies following GnRH agonist triggering (the freeze-all approach and a fresh embryo transfer attempt using a low-dose of hCG for intensified luteal phase support) are safer alternatives when compared with conventional hCG triggering with similar pregnancy outcomes. However, these two strategies have never been compared head-to-head in an unrestricted predicted hyper-responder population. This study included women with an excessive response to ovarian stimulation (≥18 follicles measuring ≥11 mm) undergoing IVF/ICSI in a GnRH antagonist suppressed cycle between 2014 and 2017. Our primary outcome was clinical pregnancy at 7 weeks after the first embryo transfer. Secondary outcomes included live birth and the development of moderate-to-severe OHSS. Following GnRH agonist triggering, women were randomized either to cryopreserve all good-quality embryos followed by a frozen embryo transfer in an subsequent artificial cycle or to perform a fresh embryo transfer with intensified luteal phase support (1500 IU hCG on the day of oocyte retrieval, plus oral estradiol 2 mg two times a day, plus 200 mg of micronized vaginal progesterone three times a day). A total of 212 patients (106 in each arm) were recruited in the study, with three patients (one in the fresh embryo transfer group and two in the freeze-all group) later withdrawing their consent to participate in the study. One patient in the freeze-all group became pregnant naturally (clinical pregnancy diagnosed 38 days after randomization) prior to the first frozen embryo transfer. The study arms did not vary significantly in terms of the number of oocytes retrieved and embryos produced/transferred. The intention to treat clinical pregnancy and live birth rates (with the latter excluding four cases lost to follow-up: one in the fresh transfer and three in the freeze-all arms, respectively) after the first embryo transfer did not vary significantly among the fresh embryo transfer and freeze-all study arms: 51/105 (48.6%) versus 57/104 (54.8%) and 41/104 (39.4%) versus 42/101 (41.6%), respectively (relative risk for clinical pregnancy 1.13, 95% CI 0.87-1.47; P = 0.41). However, moderate-to-severe OHSS occurred solely in the group that received low-dose hCG (9/105, 8.6%, 95% CI 3.2% to 13.9% vs 0/104, 95% CI 0 to 3.7, P < 0.01). The sample size calculation was based on a 19% absolute difference in terms of clinical pregnancy rates, therefore smaller differences, as observed in the trial, cannot be reliably excluded as non-significant. This study offers the first comparative analysis of two common strategies applied to women performing IVF/ICSI with a high risk to develop OHSS. While pregnancy rates did not vary significantly, a fresh embryo transfer with intensified luteal phase support may still not avoid the risk of moderate-to-severe OHSS and serious consideration should be made before recommending it as a routine first-line treatment. Future trials may allow us to confirm these findings. The authors have no conflicts of interest to disclose. No external funding was obtained for this study. ClinicalTrials.gov identifier NCT02148393. 28 May 2014. 30 May 2014.


DNA assembly using bis-peptide nucleic acids (bisPNAs).
DNA nanostructures are ordered oligonucleotide arrangements that have applications for DNA computers, crystallography, diagnostics and material sciences. Peptide nucleic acid (PNA) is a DNA/RNA mimic that offers many advantages for hybridization, but its potential for application in the field of DNA nanotechnology has yet to be thoroughly examined. We report the synthesis and characterization of tethered PNA molecules (bisPNAs) designed to assemble two individual DNA molecules through Watson-Crick base pairing. The spacer regions linking the PNAs were varied in length and contained amino acids with different electrostatic properties. We observed that bisPNAs effectively assembled oligonucleotides that were either the exact length of the PNA or that contained overhanging regions that projected outwards. In contrast, DNA assembly was much less efficient if the oligonucleotides contained overhanging regions that projected inwards. Surprisingly, the length of the spacer region between the PNA sequences did not greatly affect the efficiency of DNA assembly. Reasons for inefficient assembly of inward projecting DNA oligonucleotides include non-sequence-specific intramolecular interactions between the overhanging region of the bisPNA and steric conflicts that complicate simultaneous binding of two inward projecting strands. These results suggest that bisPNA molecules can be used for self-assembling DNA nanostructures provided that the arrangement of the hybridizing DNA oligonucleotides does not interfere with simultaneous hybridization to the bisPNA molecule.


The International Index of Erectile Function (IIEF)-A Systematic Review of Measurement Properties.
The International Index of Erectile Function (IIEF) is a patient-reported outcome measure to evaluate erectile dysfunction and other sexual problems in men. To perform a systematic review of the measurement properties of the 15-item patient-reported outcome measure (IIEF-15) and the shortened 5-item version (IIEF-5). A systematic search of scientific literature up to April 2018 was performed. Data were extracted and analyzed according to COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) guidelines for structural validity, internal consistency, reliability, measurement error, hypothesis testing for construct validity, and responsiveness. Evidence of measurement properties was categorized into sufficient, insufficient, inconsistent, or indeterminate, and quality of evidence as very high, high, moderate, or low. 40 studies were included. The evidence for criterion validity (of the Erectile Function subscale), and responsiveness of the IIEF-15 was sufficient (high quality), but inconsistent (moderate quality) for structural validity, internal consistency, construct validity, and test-retest reliability. Evidence for structural validity, test-retest reliability, construct validity, and criterion validity of the IIEF-5 was sufficient (moderate quality) but indeterminate for internal consistency, measurement error, and responsiveness. Lack of evidence for and evidence not supporting some of the measurement properties of the IIEF-15 and IIEF-5 shows the importance of further research on the validity of these questionnaires in clinical research and clinical practice. A strength of the current review is the use of predefined guidelines (COSMIN). A limitation of this review is the use of a precise rather than a sensitive search filter regarding measurement properties to identify studies to be included. The IIEF requires more research on structural validity (IIEF-15), internal consistency (IIEF-15 and IIEF-5), construct validity (IIEF-15), measurement error (IIEF-15 and IIEF-5), and responsiveness (IIEF-5). The most pressing matter for future research is determining the unidimensionality of the IIEF-5 and the exact factor structure of the IIEF-15. Neijenhuijs KI, Holtmaat K, Aaronson NK, et al. The International Index of Erectile Function (IIEF)-A Systematic Review of Measurement Properties. J Sex Med 2019;16:1078-1091.


eEF-2 kinase, another meddler in the "yin and yang" of Akt-mediated cell fate?
Eukaryotic elongation factor-2 (eEF-2) kinase, also known as calmodulin-dependent protein kinase III, is a unique calcium/calmodulin-dependent enzyme. eEF-2 kinase can act as a negative regulator of protein synthesis and a positive regulator of autophagy under environmental or metabolic stresses. Akt, a key downstream effector of the PI3K signaling pathway that regulates cell survival and proliferation, is an attractive therapeutic target for anticancer treatment. Akt inhibition leads to activation of both apoptosis, type I programmed cell death and autophagy, a cellular degradation process via lysosomal machinery (also termed type II programmed cell death). However, the underlying mechanisms that dictate functional relationship between autophagy and apoptosis in response to Akt inhibition remain to be delineated. Our recent study demonstrated that inhibition of eEF-2 kinase can suppress autophagy but promote apoptosis in tumor cells subjected to Akt inhibition, indicating a role of eEF-2 kinase as a controller in the crosstalk between autophagy and apoptosis. Furthermore, inhibition of eEF-2 kinase can reinforce the efficacy of a novel Akt inhibitor, MK-2206, against human glioma. These findings may help optimize the use of Akt inhibitors in the treatment of cancer and other diseases.


Female giant panda (Ailuropoda melanoleuca) chirps advertise the caller's fertile phase.
Although female mammal vocal behaviour is known to advertise fertility, to date, no non-human mammal study has shown that the acoustic structure of female calls varies significantly around their fertile period. Here, we used a combination of hormone measurements and acoustic analyses to determine whether female giant panda chirps have the potential to signal the caller's precise oestrous stage (fertile versus pre-fertile). We then used playback experiments to examine the response of male giant pandas to female chirps produced during fertile versus pre-fertile phases of the caller's reproductive cycle. Our results show that the acoustic structure of female giant panda chirps differs between fertile and pre-fertile callers and that male giant pandas can perceive differences in female chirps that allow them to determine the exact timing of the female's fertile phase. These findings indicate that male giant pandas could use vocal cues to preferentially associate and copulate with females at the optimum time for insemination and reveal the likely importance of female vocal signals for coordinating reproductive efforts in this critically endangered species.


Psychophysical investigations of the temporal modulation sensitivity function in amblyopia: uniform field flicker.
The temporal modulation sensitivity function was studied in eight strabismic and/or anisometropic amblyopes and in two subjects with amblyopia resulting from monocular stimulus deprivation (cataract and ptosis). Half of the strabismic/anisometropic amblyopes showed a reduction in the sensitivity of the amblyopic eye. These differences were more marked at low and middle temporal frequencies. Two strabismic/anisometropic observers showed little difference in sensitivity between the two eyes, but in the remaining two observers the amblyopic eye was slightly more sensitive to low-frequency modulation than was the nonamblyopic eye. Four of the strabismic/anisometropic amblyopes also showed a small (5% to 10%) but statistically significant difference in the critical fusion frequency, with the sensitivity of the amblyopic eye being reduced. These differences in sensitivity were not related to the visual acuity loss. However, all differences in sensitivity between the two eyes were reduced or eliminated when the mean luminance of the test field was decreased by 2 log units form 32 to 0.32 cd/m2, or with suprathreshold stimulation. One of the two stimulus-deprivation amblyopes showed a decrease in sensitivity to low and middle temporal modulation frequencies similar to that found in four of in sensitivity to low and middle temporal modulation frequencies similar to similar to that found in four of the strabismic/anisometropic amblyopes, and the other observer showed a marked decrease in sensitivity at all temporal frequencies. The losses in temporal resolution of the amblyopic eye in both strabismic/anisometropic and stimulus-deprivation amblyopia are small in comparison with those reported in the spatial domain.


A Robust Quasi-Quantum Walks-Based Steganography Protocol for Secure Transmission of Images on Cloud-Based E-healthcare Platforms.
Traditionally, tamper-proof steganography involves using efficient protocols to encrypt the stego cover image and/or hidden message prior to embedding it into the carrier object. However, as the inevitable transition to the quantum computing paradigm beckons, its immense computing power will be exploited to violate even the best non-quantum, i.e., classical, stego protocol. On its part, quantum walks can be tailored to utilise their astounding 'quantumness' to propagate nonlinear chaotic behaviours as well as its sufficient sensitivity to alterations in primary key parameters both important properties for efficient information security. Our study explores using a classical (i.e., quantum-inspired) rendition of the controlled alternate quantum walks (i.e., CAQWs) model to fabricate a robust image steganography protocol for cloud-based E-healthcare platforms by locating content that overlays the secret (or hidden) bits. The design employed in our technique precludes the need for pre and/or post encryption of the carrier and secret images. Furthermore, our design simplifies the process to extract the confidential (hidden) information since only the stego image and primary states to run the CAQWs are required. We validate our proposed protocol on a dataset of medical images, which exhibited remarkable outcomes in terms of their security, good visual quality, high resistance to data loss attacks, high embedding capacity, etc., making the proposed scheme a veritable strategy for efficient medical image steganography.


Diagnostic and prognostic utility of TROP-2, SLP-2, and CXCL12 expression in papillary thyroid carcinoma.
Papillary thyroid carcinoma (PTC) is the most frequent thyroid malignancy. Histopathological examination is widely accepted as the gold standard test for the diagnosis of PTC. However, the histopathological examination sometimes can't differentiate PTC from other thyroid diseases. Differentiating PTC from other thyroid diseases is essential for a therapeutic approach and prognosis. The current study was performed to investigate the utility of TROP-2, SPL-2, and CXCL12 mRNA and protein expression in discriminating PTC from other thyroid diseases that mimic PTC. The current study was performed on 75 cases of surgically resected thyroid glands. The cases were distributed in two groups: the PTC group and the non-PTC group. The PTC group consisted of 35 cases (25 patients of the classic PTC variant and 10 patients of the PTC follicular variant). The non-PTC group consisted of 40 cases (10 cases were multinodular goiter, 5 cases were Graves' disease, 5 cases were Hashimoto thyroiditis, 15 patients were follicular adenoma (FA) and 5 cases were follicular carcinoma). TROP-2, SPL-2, and CXCL12 mRNA expression were estimated by qRT-PCR, and protein expression was estimated by immunohistochemistry. There were upregulated TROP-2, SPL-2, and CXCL12 mRNA and protein expressions in PTC compared to non-PTC (P< 0.001, for each). There was a statistically significant upregulation in the mRNA expression of the three genes among PTC cases with larger tumor sizes (P< 0.001, for each), those with tumor stages III and IV (P= 0.008, 0.002 and < 0.001 respectively), and those with LN metastasis (P< 0.001, for each). Moreover, there was a statistically significant upregulation in CXCL-12 gene expression among PTC cases with extra-thyroid extension (P< 0.001). mRNA expression of TROP-2, SPL-2, and CXCL12 among PTC cases increased in larger tumor size, tumor stages III and IV, and LN metastasis. Moreover, there was an increase in CXCL-12 gene expression among PTC cases with extra-thyroid extension. Thus, TROP-2, SPL-2, and CXCL12 expressions could be possible diagnostic and prognostic markers in PTC.


The relationships between family primary caregiver characteristics and satisfaction with hospice care, quality of life, and burden.
To examine relationships between family primary caregiver characteristics and satisfaction with hospice care, quality of life (QOL), and burden. Exploratory, quantitative. Five hospice organizations in Eastern Washington and Northern Idaho. 44 primary caregivers of patients enrolled in hospice for more than two weeks. Telephone surveys measured primary caregivers' satisfaction with hospice care, QOL, and burden. Demographic, personal, and situational characteristics were compiled. Multivariate regression techniques were used to identify caregiver characteristics that explained the most variation in satisfaction with hospice care, QOL, and burden. Satisfaction with hospice care, QOL, and burden. Being retired, being a wife or daughter, patient diagnosis, per diem rate paid to the hospice organization, county population and density, length of time in hospice, and length of time as a caregiver were significantly related to satisfaction with hospice care, QOL, and burden. Age, sex, education level, occupation, income, patient functional status, and social support were not related to these variables. Although caregivers reported satisfaction with hospice care, the caregiver role negatively affects their QOL and they are burdened. "At-risk" caregivers are still working, have been providing care for a long time, and live in a rural locale. These findings create a profile of "typical" and "at-risk" primary caregivers and support the role of hospice to care for family caregivers.


Body mass index, waist/hip ratio, and coronary heart disease incidence in African Americans and whites. Atherosclerosis Risk in Communities Study Investigators.
To study the relation of the amount and distribution of body fat with incident coronary heart disease in two ethnic groups, the authors analyzed prospective data from the Atherosclerosis Risk in Communities Study. Among 14,040 participants aged 45-64 years and free of coronary disease at baseline in 1987-1989, we identified 398 events through 1994, an average of 6.2 years of follow-up. Among African-American women, the multivariable-adjusted relative risks of coronary heart disease across quartiles of body mass index were 1.0, 1.91. 1.54, and 2.15 (p for trend=0.27), and those for waist/hip ratio were 1.0, 2.07, 2.33, and 4.22 (p for trend=0.02). Among African-American men, these respective relative risks were 1.0, 1.03, 0.83, and 1.20 (p for trend=0.76) for body mass index and 1.0, 1.08, 1.87, and 1.68 (p for trend=0.06) for waist/hip ratio. Relative risks for whites were generally similar to those for African Americans. Relative risks were stronger for never smokers than for the overall cohort. Unlike some previous studies, our results suggest that Africa Americans, like whites, are not spared from the coronary heart disease risks accompanying obesity.


Reducing isozyme competition increases target fatty acid accumulation in seed triacylglycerols of transgenic Arabidopsis.
One goal of green chemistry is the production of industrially useful fatty acids (FAs) in crop plants. We focus on hydroxy fatty acids (HFAs) and conjugated polyenoic FAs (α-eleostearic acids [ESAs]) using Arabidopsis (Arabidopsis thaliana) as a model. These FAs are found naturally in seed oils of castor (Ricinus communis) and tung tree (Vernicia fordii), respectively, and used for the production of lubricants, nylon, and paints. Transgenic oils typically contain less target FA than that produced in the source species. We hypothesized that competition between endogenous and transgenic isozymes for substrates limits accumulation of unique FAs in Arabidopsis seeds. This hypothesis was tested by introducing a mutation in Arabidopsis diacylglycerol acyltransferase1 (AtDGAT1) in a line expressing castor FA hydroxylase and acyl-Coenzyme A:RcDGAT2 in its seeds. This led to a 17% increase in the proportion of HFA in seed oil. Expression of castor phospholipid:diacylglycerol acyltransferase 1A in this line increased the proportion of HFA by an additional 12%. To determine if our observations are more widely applicable, we investigated if isozyme competition influenced production of ESA. Expression of tung tree FA conjugase/desaturase in Arabidopsis produced approximately 7.5% ESA in seed lipids. Coexpression of VfDGAT2 increased ESA levels to approximately 11%. Overexpression of VfDGAT2 combined with suppression of AtDGAT1 increased ESA accumulation to 14% to 15%. Our results indicate that isozyme competition is a limiting factor in the engineering of unusual FAs in heterologous plant systems and that reduction of competition through mutation and RNA suppression may be a useful component of seed metabolic engineering strategies.


Caveolin-1 mediates Fas-BID signaling in hyperoxia-induced apoptosis.
Fas-mediated apoptosis is a crucial cellular event. Fas, the Fas-associated death domain, and caspase 8 form the death-inducing signaling complex (DISC). Activated caspase 8 mediates the extrinsic pathways and cleaves cytosolic BID. Truncated BID (tBID) translocates to the mitochondria, facilitates the release of cytochrome c, and activates the intrinsic pathways. However, the mechanism causing these DISC components to aggregate and form the complex remains unclear. We found that Cav-1 regulated Fas signaling and mediated the communication between extrinsic and intrinsic pathways. Shortly after hyperoxia (4 h), the colocalization and interaction of Cav-1 and Fas increased, followed by Fas multimer and DISC formation. Deletion of Cav-1 (Cav-1-/-) disrupted DISC formation. Further, Cav-1 interacted with BID. Mutation of Cav-1 Y14 tyrosine to phenylalanine (Y14F) disrupted the hyperoxia-induced interaction between BID and Cav-1 and subsequently yielded a decreased level of tBID and resistance to hyperoxia-induced apoptosis. The reactive oxygen species (ROS) scavenger N-acetylcysteine decreased the Cav-1-Fas interaction. Deletion of glutathione peroxidase-2 using siRNA aggravated the BID-Cav-1 interaction and tBID formation. Taken together, these results indicate that Cav-1 regulates hyperoxia/ROS-induced apoptosis through interactions with Fas and BID, probably via Fas palmitoylation and Cav-1 Y14 phosphorylation, respectively.


The Netherlands Twin Register: Longitudinal Research Based on Twin and Twin-Family Designs.
The Netherlands Twin Register (NTR) is a national register in which twins, multiples and their parents, siblings, spouses and other family members participate. Here we describe the NTR resources that were created from more than 30 years of data collections; the development and maintenance of the newly developed database systems, and the possibilities these resources create for future research. Since the early 1980s, the NTR has enrolled around 120,000 twins and a roughly equal number of their relatives. The majority of twin families have participated in survey studies, and subsamples took part in biomaterial collection (e.g., DNA) and dedicated projects, for example, for neuropsychological, biomarker and behavioral traits. The recruitment into the NTR is all inclusive without any restrictions on enrollment. These resources - the longitudinal phenotyping, the extended pedigree structures and the multigeneration genotyping - allow for future twin-family research that will contribute to gene discovery, causality modeling, and studies of genetic and cultural inheritance.


Structural analysis of Arabidopsis thaliana chromosome 5. VII. Sequence features of the regions of 1,013,767 bp covered by sixteen physically assigned P1 and TAC clones.
Sixteen P1 and TAC clones assigned to Arabidopsis thaliana chromosome 5 were sequenced, and their sequence features were analyzed using various computer programs. The total length of the sequences determined was 1,013,767 bp. Together with the nucleotide sequences of 109 clones previously reported, the regions of chromosome 5 sequenced so far now total 9,072,622 bp, which presumably covers approximately one-third of the chromosome. A similarity search against the reported gene sequences predicted the presence of a total of 225 protein-coding genes and/or gene segments in the newly sequenced regions, indicating an average gene density of one gene per 4.5 kb. Introns were identified in 72.4% of the potential protein genes for which the entire gene structure was predicted, and the average number per gene and the average length of the introns were 3.3 and 163 bp, respectively. These sequence features are essentially identical to those in the previously reported sequences. The sequence data and gene information are available on the World Wide Web database KAOS (Kazusa Arabidopsis data Opening Site) at http://www.kazusa.or.jp/arabi/.


Tension of knotted surgical sutures shows tissue specific rapid loss in a rodent model.
Every surgical suture compresses the enclosed tissue with a tension that depends from the knotting force and the resistance of the tissue. The aim of this study was to identify the dynamic change of applied suture tension with regard to the tissue specific cutting reaction. In rabbits we placed single polypropylene sutures (3/0) in skin, muscle, liver, stomach and small intestine. Six measurements for each single organ were determined by tension sensors for 60 minutes. We collected tissue specimens to analyse the connective tissue stability by measuring the collagen/protein content. We identified three phases in the process of suture loosening. The initial rapid loss of the first phase lasts only one minute. It can be regarded as cutting through damage of the tissue. The percentage of lost tension is closely related to the collagen content of the tissue (r = -0.424; p = 0.016). The second phase is characterized by a slower decrease of suture tension, reflecting a tissue specific plastic deformation. Phase 3 is characterized by a plateau representing the remaining structural stability of the tissue. The ratio of remaining tension to initial tension of phase 1 is closely related to the collagen content of the tissue (r = 0.392; p = 0.026). Knotted non-elastic monofilament sutures rapidly loose tension. The initial phase of high tension may be narrowed by reduction of the surgeons' initial force of the sutures' elasticity to those of the tissue. Further studies have to confirm, whether reduced tissue compression and less local damage permits improved wound healing.


Quantification of fecal carbohydrates by near-infrared reflectance analysis.
Established methods for quantitative analysis of fecal carbohydrates (CHO) are time consuming, require unpopular sample handling, and are therefore rarely performed. The aim of this study was to evaluate the efficiency, validity, and practicability of near-infrared reflectance analysis (NIRA), compared with standard methods for measurement of fecal CHO. Excretion of fecal CHO was cross- validated spectrophotometrically with the anthrone method. Fecal CHO concentrations ranged form 2.7 to 24.5 g/kg wet weight. Methods comparison showed linear regression over the entire range of diagnostic relevance with a correlation coefficient of 0.869 (S(y/x) +/- 0.31). Repeated measurements from the same stool collections obviated the need for homogenization. These results indicate that NIRA may be a new, reliable, and accurate test in the diagnosis of CHO malabsorption.


Hierarchical structures and complexities of parallel isometric languages.
The relationship between parallel isometric array languages and sequential isometric array languages is examined. Their hierarchical structures are investigated and a hierarchy is established by introducing parallel context-free array languages (PCFAL), derivation bounded array languages (DBAL), linear array languages (LAL), and extended regular afray languages (ERAL). It is interesting to find that some fundamental aspects that hold in one-dimensional string languages do not hold in their two-dimensional counterparts. Some parsing techniques are also explored. It is shown that while parallel parsing grammars may be simpler to write and parallel processing usually takes less time than sequential ones, the nature of parallel parsing is very complicated. Finally, several future research topics concerned with parallel isometric array languages including their complexities, hierarchical structures, and application to pattern recognition are discussed.


Effects of exogenous heparin on the vasculogenesis of the chorioallantoic membrane.
On the basis of the well-known stimulating role played by heparin on blood vessel neoformation in the chorioallantoic membrane (CAM), demonstrated by means of slow-release polymers soaked in this substance and placed on CAMs, a new technique of introducing heparin directly into the allantoic sac is proposed. According to observations carried out in ovo and after fixation, morphological modifications are demonstrated in the developing vasculature of the heparin-treated CAMs which, compared with the control CAMs, show dilated and sinuous arterial and venous branches, denser and irregular capillary networks, and a high number of vascular primordia. The results, confirming that heparin is involved in angiogenesis, indicate the suitability of the technique and suggest that the effects of this substance could be enhanced by growth factors released by CAM tissues rapidly growing up from the 5th to the 7th incubation day.


Establishment of a novel xenograft model for human uterine leiomyoma in immunodeficient mice.
Uterine leiomyomas are the most common gynecological benign tumor and greatly affect reproductive health and wellbeing, but the pathophysiology and epidemiology of uterine leiomyoma are poorly understood. One of the major reasons for the slow progress in leiomyoma research is the lack of a good in vivo model system. We therefore aimed to develop a novel model by transplanting human uterine leiomyoma xenografts in an immunodeficient mouse strain (NOD/SCID/gammac-null: NOG). Human uterine leiomyoma tissues were cut into small pieces and inserted subcutaneously into the right and left flanks of NOG mice. Estrogen supplementation was needed to maintain the features of uterine leiomyoma in xenografted tissues. After 4 weeks or 8 weeks of transplantation, xenografted tissues were harvested and analyzed regarding tissue morphology, collagen content, and proliferation and apoptosis of uterine leiomyoma smooth muscle cells. The xenografts that were harvested after 4 weeks and 8 weeks retained the histological architecture of original uterine leiomyoma tissue both in cellular and collagen components. The expression profiles of key markers of uterine leiomyoma were also maintained, including estrogen receptor, progesterone receptor, and alpha-smooth muscle actin, as judged by immunohistochemical staining. The proportion of proliferating cells was significantly increased (1.5-fold) in the xenografts after 8 weeks of transplantation, whereas that of the apoptotic cells remained unchanged. Importantly, the reproducible results were obtained with the tumor tissues derived from six patients. The present in vivo model may provide a useful tool for development of novel therapeutic strategies for uterine leiomyoma.


Key Findings and Comparisons From Analogous Case-Cluster Studies for Dengue Virus Infection Conducted in Machala, Ecuador, and Kamphaeng Phet, Thailand.
Dengue viruses (DENV) pose a significant and increasing threat to human health across broad regions of the globe. Currently, prevention, control, and treatment strategies are limited. Promising interventions are on the horizon, including multiple vaccine candidates under development and a renewed and innovative focus on controlling the vector, Aedes aegypti. However, significant gaps persist in our understanding of the similarities and differences in DENV epidemiology across regions of potential implementation and evaluation. In this manuscript, we highlight and compare findings from two analogous cluster-based studies for DENV transmission and pathogenesis conducted in Thailand and Ecuador to identify key features and questions for further pursuit. Despite a remarkably similar incidence of DENV infection among enrolled neighborhood contacts at the two sites, we note a higher occurrence of secondary infection and severe illness in Thailand compared to Ecuador. A higher force of infection in Thailand, defined as the incidence of infection among susceptible individuals, is suggested by the higher number of captured Aedes mosquitoes per household, the increasing proportion of asymptomatic infections with advancing age, and the high proportion of infections identified as secondary-type infections by serology. These observations should be confirmed in long-term, parallel prospective cohort studies conducted across regions, which would advantageously permit characterization of baseline immune status (susceptibility) and contemporaneous assessment of risks and risk factors for dengue illness.


New application of tiplaxtinin as an effective FtsZ-targeting chemotype for an antimicrobial study.
The filamenting temperature-sensitive mutant Z (FtsZ) protein is generally recognized as a promising antimicrobial drug target. In the present study, a small organic molecule (tiplaxtinin) was identified for the first time as an excellent cell division inhibitor by using a cell-based screening approach from a library with 250 compounds. Tiplaxtinin possesses potent antibacterial activity against Gram-positive pathogens. Both in vitro and in vivo results reveal that the compound is able to disrupt dynamic assembly of FtsZ and Z-ring formation effectively through the mechanism of stimulating FtsZ polymerization and impairing GTPase activity.


Inequity in maternal health care service utilization in Gujarat: analyses of district-level health survey data.
Two decades after the launch of the Safe Motherhood campaign, India still accounts for at least a quarter of maternal death globally. Gujarat is one of the most economically developed states of India, but progress in the social sector has not been commensurate with economic growth. The purpose of this study was to use district-level data to gain a better understanding of equity in access to maternal health care and to draw the attention of the policy planers to monitor equity in maternal care. Secondary data analyses were performed among 7,534 ever-married women who delivered since January 2004 in the District Level Household and Facility Survey (DLHS-3) carried out during 2007-2008 in Gujarat, India. Based on the conceptual framework designed by the Commission on the Social Determinants of Health, associations were assessed between three outcomes - Institutional delivery, antenatal care (ANC), and use of modern contraception - and selected intermediary and structural determinants of health using multiple logistic regression. Inequities in maternal health care utilization persist in Gujarat. Structural determinants like caste group, wealth, and education were all significantly associated with access to the minimum three antenatal care visits, institutional deliveries, and use of any modern method of contraceptive. There is a significant relationship between being poor and access to less utilization of ANC services independent of caste category or residence. Poverty is the most important determinant of non-use of maternal health services in Gujarat. In addition, social position (i.e. caste) has a strong independent effect on maternal health service use. More focused and targeted efforts towards these disadvantaged groups needs to be taken at policy level in order to achieve targets and goals laid out as per the MDGs. In particular, the Government of Gujarat should invest more in basic education and infrastructural development to begin to remove the structural causes of non-use of maternal health services.


[Effect of spironolactone in patients with heart failure and preserved left ventricular function - TOPCAT study].
The TOPCAT study followed the effect of spironolactone on a chronic heart failure with a preserved left ventricular ejection fraction. The study did not find any impact on the primary goal of the study, i.e. a combination of cardiovascular mortality, managed cardiac arrest or hospitalization rate for heart failure treatment. The only finding of the study was the decrease in hospitalization rates for a heart failure. A post hoc analysis, however, identified a significant difference between patients from the Americas (USA, Canada, Argentina, Brazil) on the one hand and those from Russia/Georgia on the other. The differences were rather striking. Whereas the former manifested a significant reduction in the incidence of the primary goal, i.e. cardiovascular mortality and hospitalization with heart failure through treatment with spironolactone, the incidence of all clinical events was considerably lower in Russia/Georgia and no impact of spironolactone at all on any of the goals was identified regarding the patients from Russia/Georgia. This post hoc analysis therefore suggested a possible therapeutic effect of spironolactone in the Americas, as well as in the populations of similar characteristics. Most studies accept a definition only based on the heart failure with a preserved left ventricular ejection fraction. The study points to the need to further elaborate this definition which also has to consider changes of the left ventricular diastolic function when defining a diastolic heart failure.


Characterization of signal bias at the GLP-1 receptor induced by backbone modification of GLP-1.
The glucagon-like peptide-1 receptor (GLP-1R) is a class B G protein-coupled receptor that is a major therapeutic target for the treatment of type 2 diabetes. Activation of this receptor promotes insulin secretion and blood glucose regulation. The GLP-1R can initiate signaling through several intracellular pathways upon activation by GLP-1. GLP-1R ligands that preferentially stimulate subsets among the natural signaling pathways ("biased agonists") could be useful as tools for elucidating the consequences of specific pathways and might engender therapeutic agents with tailored effects. Using HEK-293 cells recombinantly expressing human GLP-1R, we have previously reported that backbone modification of GLP-1, via replacement of selected α-amino acid residues with β-amino acid residues, generates GLP-1 analogues with distinctive preferences for promoting G protein activation versus β-arrestin recruitment. Here, we have explored the influence of cell background across these two parameters and expanded our analysis to include affinity and other key signaling pathways (intracellular calcium mobilization and ERK phosphorylation) using recombinant human GLP-1R expressed in a CHO cell background, which has been used extensively to demonstrate biased agonism of GLP-1R ligands. The new data indicate that α/β-peptide analogues of GLP-1 exhibit a range of distinct bias profiles relative to GLP-1 and that broad assessment of signaling endpoints is required to reveal the spectrum of behavior of modified peptides. These results support the view that backbone modification via α→β amino acid replacement can enable rapid discovery of peptide hormone analogues that display substantial signal bias at a cognate GPCR.


Real-world efficacy of adjuvant single-agent intravesical gemcitabine for non-muscle invasive bladder cancer.
Failure, intolerance, or shortage of bacillus Calmette-Guerin (BCG) treatment for patients with high-risk (HR) non-muscle invasive bladder cancer (NMIBC) leave many facing the prospect of radical cystectomy (RC). However, despite the lack of large-scale randomized controlled studies with single-agent intravesical gemcitabine (Gem), it has emerged as a popular salvage agent after BCG failure or even a treatment alternative to BCG. 1. Characterization of treatment regimen details pertaining to single-agent intravesical adjuvant Gem use among disease states of NMIBC characterized by risk and BCG exposure. 2. Comparison of safety and efficacy of Gem according to risk category, type of tumor (papillary vs. carcinoma in situ (CIS)), and tumor grades. Two randomized studies in early BCG failure disease demonstrate that single-agent Gem has superior efficacy versus repeated BCG therapy or mitomycin C. Studies enrolling patients with predominantly papillary disease without CIS, intermediate-risk (IR) disease, and less BCG exposure appear to derive the highest benefits from adjuvant Gem in terms of recurrence and progression. However, studies with cohorts enriched for a predominance of CIS, HR disease and/or more extensive BCG failure have poorer 2-year recurrence free survival and a somewhat higher risk of progression and RC.


Not everything is black and white: color and behavioral variation reveal a continuum between cryptic and aposematic strategies in a polymorphic poison frog.
Aposematism and crypsis are often viewed as two extremes of a continuum of visual conspicuousness to predators. Theory predicts that behavioral and coloration conspicuousness should vary in tandem along the conspicuousness spectrum for antipredator strategies to be effective. Here we used visual modeling of contrast and behavioral observations to examine the conspicuousness of four populations of the granular poison frog, Oophaga granulifera, which exhibits almost continuous variation in dorsal color. The patterns of geographic variation in color, visual contrast, and behavior support a gradient of overall conspicuousness along the distribution of O. granulifera. Red and green populations, at the extremes of the color distribution, differ in all elements of color, contrast, and behavior, strongly reflecting aposematic and cryptic strategies. However, there is no smooth cline in any elements of behavior or coloration between the two extremes. Instead populations of intermediate colors attain intermediate conspicuousness by displaying different combinations of aposematic and cryptic traits. We argue that coloration divergence among populations may be linked to the evolution of a gradient of strategies to balance the costs of detection by predators and the benefits of learned aversion.


Loss channels in triplet-triplet annihilation photon upconversion: importance of annihilator singlet and triplet surface shapes.
Triplet-triplet annihilation photon upconversion (TTA-UC) can, through a number of energy transfer processes, efficiently combine two low frequency photons into one photon of higher frequency. TTA-UC systems consist of one absorbing species (the sensitizer) and one emitting species (the annihilator). Herein, we show that the structurally similar annihilators, 9,10-diphenylanthracene (DPA, 1), 9-(4-phenylethynyl)-10-phenylanthracene (2) and 9,10-bis(phenylethynyl)anthracene (BPEA, 3) have very different upconversion efficiencies, 15.2 ± 2.8%, 15.9 ± 1.3% and 1.6 ± 0.8%, respectively (of a maximum of 50%). We show that these results can be understood in terms of a loss channel, previously unaccounted for, originating from the difference between the BPEA singlet and triplet surface shapes. The difference between the two surfaces results in a fraction of the triplet state population having geometries not energetically capable of forming the first singlet excited state. This is supported by TD-DFT calculations of the annihilator excited state surfaces as a function of phenyl group rotation. We thereby highlight that the commonly used "spin-statistical factor" should be used with caution when explaining TTA-efficiencies. Furthermore, we show that the precious metal free zinc octaethylporphyrin (ZnOEP) can be used for efficient sensitization and that the upconversion quantum yield is maximized when sensitizer-annihilator spectral overlap is minimized (ZnOEP with 2).


Metabolic effects of resistance or high-intensity interval training among glycemic control-nonresponsive children with insulin resistance.
Little evidence exists on which variables of body composition or muscular strength mediates more glucose control improvements taking into account inter-individual metabolic variability to different modes of exercise training. We examined 'mediators' to the effects of 6-weeks of resistance training (RT) or high-intensity interval training (HIT) on glucose control parameters in physically inactive schoolchildren with insulin resistance (IR). Second, we also determined both training-induce changes and the prevalence of responders (R) and non-responders (NR) to decrease the IR level. Fifty-six physically inactive children diagnosed with IR followed a RT or supervised HIT program for 6 weeks. Participants were classified based on ΔHOMA-IR into glycemic control R (decrease in homeostasis model assessment-IR (HOMA-IR) <3.0 after intervention) and NRs (no changes or values HOMA-IR⩾3.0 after intervention). The primary outcome was HOMA-IR associated with their mediators; second, the training-induced changes to glucose control parameters; and third the report of R and NR to improve body composition, cardiovascular, metabolic and performance variables. Mediation analysis revealed that improvements (decreases) in abdominal fat by the waist circumference can explain more the effects (decreases) of HOMA-IR in physically inactive schoolchildren under RT or HIT regimes. The same analysis showed that increased one-maximum repetition leg-extension was correlated with the change in HOMA-IR (β=-0.058; P=0.049). Furthermore, a change in the waist circumference fully mediated the dose-response relationship between changes in the leg-extension strength and HOMA-IR (β'=-0.004; P=0.178). RT or HIT were associated with significant improvements in body composition, muscular strength, blood pressure and cardiometabolic parameters irrespective of improvement in glycemic control response. Both glucose control RT-R and HIT-R (respectively), had significant improvements in mean HOMA-IR, mean muscular strength leg-extension and mean measures of adiposity. The improvements in the lower body strength and the decreases in waist circumference can explain more the effects of the improvements in glucose control of IR schoolchildren in R group after 6 weeks of RT or HIT, showing both regimes similar effects on body composition or muscular strength independent of interindividual metabolic response variability.


[Scalp neuralgia and headache elicited by cranial superficial anatomical causes: supraorbital neuralgia, occipital neuralgia, and post-craniotomy headache].
Most scalp neuralgias are supraorbital or occipital. Although they have been considered idiopathic, recent studies revealed that some were attributable to mechanical irritation with the peripheral nerve of the scalp by superficial anatomical cranial structures. Supraorbital neuralgia involves entrapment of the supraorbital nerve by the facial muscle, and occipital neuralgia involves entrapment of occipital nerves, mainly the greater occipital nerve, by the semispinalis capitis muscle. Contact between the occipital artery and the greater occipital nerve in the scalp may also be causative. Decompression surgery to address these neuralgias has been reported. As headache after craniotomy is the result of iatrogenic injury to the peripheral nerve of the scalp, post-craniotomy headache should be considered as a differential diagnosis.


Type II Epithelial-Mesenchymal Transition Upregulates Protein N-Glycosylation To Maintain Proteostasis and Extracellular Matrix Production.
Type II epithelial-mesenchymal transition (EMT) plays a vital role in airway injury, repair, and remodeling. Triggered by growth factors, such as transforming growth factor beta (TGFβ), EMT induced a biological process that converts epithelial cells into secretory mesenchymal cells with a substantially increased production of extracellular matrix (ECM) proteins. Epithelial cells are not professional secretory cells and produce few ECM proteins under normal conditions. The molecular mechanism underlying the transformation of the protein factory and secretory machinery during EMT is significant because ECM secretion is central to the pathogenesis of airway remodeling. Here we report that type II EMT upregulates the protein N-glycosylation of ECMs. The mechanism study reveals that the substantial increase in synthesis of ECM proteins in EMT activates the inositol-requiring protein 1 (IRE1α)-X-box-binding protein 1 (XBP1) axis of the unfolded protein response (UPR) coupled to the hexosamine biosynthesis pathway (HBP). These two pathways coordinately up-regulate the protein N-glycosylation of ECM proteins and increase ER folding capacity and ER-associated degradation (ERAD), which improve ER protein homeostasis and protect transitioned cells from proteotoxicity. Inhibition of the alternative splicing of XBP1 or protein N-glycosylation blocks ECM protein secretion, indicating the XBP1-HBP plays a prominent role in regulating the secretion of ECM proteins in the mesenchymal transition. Our data suggest that the activation of XBP1-HBP pathways and elevation of protein N-glycosylation is an adaptive response to maintain protein quality control and facilitate the secretion of ECM proteins during the mesenchymal transition. The components of the XBP1-HBP pathways may be therapeutic targets to prevent airway remodeling.


Treatment of Super-Refractory Status Epilepticus: A Review.
Super-refractory status epilepticus (SRSE) presents management challenges due to the absence of randomized controlled trials and a plethora of potential medical therapies. The literature on treatment options for SRSE reports variable success and quality of evidence. This review is a sequel to the 2020 American Epilepsy Society (AES) comprehensive review of the treatment of convulsive refractory status epilepticus (RSE). We sought to determine the effectiveness of treatment options for SRSE. We performed a structured literature search (MEDLINE, Embase, CENTRAL, CINAHL) for studies on reported treatments of SRSE. We excluded antiseizure medications (ASMs) covered in the 2016 AES guideline on the treatment of established SE and the convulsive RSE comprehensive review of the 2020 AES. Literature was reviewed on the effectiveness of vagus nerve stimulation, ketogenic diet (KD), lidocaine, inhalation anesthetics, brain surgery, therapeutic hypothermia, perampanel, pregabalin (PGB), and topiramate in the treatment of SRSE. Two authors reviewed each therapeutic intervention. We graded the level of the evidence according to the 2017 classification scheme of the American Academy of Neurology. For SRSE (level U; 39 class IV studies total), insufficient evidence exists to support that perampanel, PGB, lidocaine, or acute vagus nerve stimulation (VNS) is effective. For children and adults with SRSE, insufficient evidence exists to support that the KD is effective (level U; 5 class IV studies). For adults with SRSE, insufficient evidence exists that brain surgery is effective (level U, 7 class IV studies). For adults with SRSE insufficient, evidence exists that therapeutic hypothermia is effective (level C, 1 class II and 4 class IV studies). For neonates with hypoxic-ischemic encephalopathy, insufficient evidence exists that therapeutic hypothermia reduces seizure burden (level U; 1 class IV study). For adults with SRSE, insufficient evidence exists that inhalation anesthetics are effective (level U, 1 class IV study) and that there is a potential risk of neurotoxicity. For patients with SRSE insufficient, evidence exists that any of the ASMs reviewed, inhalational anesthetics, ketogenic diet, acute VNS, brain surgery, and therapeutic hypothermia are effective treatments. Data supporting the use of these treatments for SRSE are scarce and limited mainly to small case series and case reports and are confounded by differences in patients' population, and comedications, among other factors.


Corticomotor Excitability Effects of Peripheral Nerve Electrical Stimulation to the Paretic Arm in Stroke.
This study aimed to examine the corticomotor excitability changes after peripheral nerve electrical stimulation (PNS) on the stroke-impaired arm. This randomized cross-over study included 32 subjects with chronic stroke. They received a 1-hr PNS or placebo PNS by random order to the ulnar and radial nerves of the paretic arm in separate sessions. The primary outcome was excitability of the corticospinal projections for the contralateral first dorsal interosseous hand muscle in terms of slope of the recruitment curve, peak motor-evoked potential amplitude, and duration of the cortical silent period, measured with transcranial magnetic stimulation on both cerebral hemispheres. Seventeen of the subjects were measured for their paretic hand dexterity (using Purdue Pegboard Test) and pinch strength as secondary outcomes. Peripheral nerve electrical stimulation, but not placebo PNS, increased recruitment curve slope and peak motor-evoked potential amplitude in both the lesioned and nonlesioned hemispheres and lengthened the cortical silent period duration in the nonlesioned hemisphere. Peripheral nerve electrical stimulation significantly improved hand dexterity scores compared with placebo PNS. Pinch strength was not changed by the interventions. A 1-hr PNS alone to the paretic arm could increase corticomotor excitability in both hemispheres, together with hand dexterity improvement in people presented with mild-to-moderate motor impairment in the paretic upper limb after stroke. Complete the self-assessment activity and evaluation online at http://www.physiatry.org/JournalCME CME OBJECTIVES: Upon completion of this article, the readers should be able to: (1) Understand the effects of a stroke on the corticomotor excitability of the lesioned and intact hemisphere; (2) Describe the effect of peripheral nerve electrical stimulation on the lesioned and intact hemisphere corticomotor excitability following stroke; and (3) Understand the effect of peripheral nerve electrical stimulation on dexterity of the paretic hand following stroke. Advanced ACCREDITATION: The Association of Academic Physiatrists is accredited by the Accreditation Council for Continuing Medical Education to provide continuing medical education for physicians.The Association of Academic Physiatrists designates this Journal-based CME activity for a maximum of 0.75 AMA PRA Category 1 Credit(s)™. Physicians should only claim credit commensurate with the extent of their participation in the activity.


Biosynthesis and processing of legumin-like storage proteins in Lupinus angustifolius (lupin).
Synthesis, secretion and post-translational proteolysis of the storage proteins in cotyledons of Lupinus angustifolius L. (lupin) have been examined in vivo and in vitro by using a combination of pulse-chase experiments with [3H]- or [35S]-labelled amino acids, subcellular fractionation and cell-free translation from poly(A)+ (polyadenylylated) RNA or membrane-bound polyribosomes. Related polypeptides were identified by immunoprecipitation, separation on sodium dodecyl sulphate/polyacrylamide gels and fluorography. The synthesis and processing of two proteins were compared. Conglutin alpha, the 11 S protein, was found as a family of precursor polypeptides of Mr 68000-88000 when translated from poly(A)+ RNA under conditions where signal segments were not cleaved, and Mr 64000-85000 both when sequestered into the endoplasmic reticulum and when accumulated in the protein bodies. Pulse-chase labelling showed that cotyledons from early stages of development were completely incapable of further proteolysis of these precursors. Nevertheless, in the same juvenile cotyledons, the precursors of the minor storage protein conglutin gamma, two polypeptides with Mr 50000-51000, were proteolytically cleaved to mature subunits of Mr 32000 and 17000 within 2 h. Further cleavage of the precursors of conglutin alpha into families of mature subunits of Mr 21000-24000 and 42000-62000 was detected in more mature cotyledons. A model is proposed which suggests that the mature subunits are produced by a single proteolytic cleavage of each of the three major precursors of conglutin alpha and also suggests that a close similarity exists between these subunits and those of other legumin-like proteins. The enzyme responsible for this cleavage, which appears at a specific stage in the middle of cotyledonary development, seems to be an integral part of the programmed developmental sequence in these pods.


Dog ownership supports the maintenance of physical activity during poor weather in older English adults: cross-sectional results from the EPIC Norfolk cohort.
Dog ownership has been suggested to encourage physical activity in older adults and may enhance resilience to poor environmental conditions. This study investigates the role of dog ownership and walking as a means of supporting the maintenance of physical activity in older adults during periods of inclement weather. The analysis used data from the European Prospective Investigation into Cancer and Nutrition Norfolk cohort. Daily physical activity (counts per minute) and minutes of sedentary behaviour were measured using accelerometers over 7 days. Three types of environmental conditions, day length, precipitation and maximum temperature, were date matched with daily physical activity. A multilevel first-order autoregressive time-series model quantified the moderating effect of self-reported dog ownership and walking on the association between physical activity and weather factors. Among the 3123 participants, 18% reported having a dog in their households and two-thirds of dog owners walked their dogs at least once a day. Regular dog walkers were more active and less sedentary on days with the poorest conditions than non-dog owners were on the days with the best conditions. In days with the worst conditions, those who walked their dogs had 20% higher activity levels than non-dog owners and spent 30 min/day less sedentary. Those who walked dogs were consistently more physically active than those who did not regardless of environmental conditions. These large differences suggest that dog walking, where appropriate, can be a component of interventions to support physical activity in older adults.


Randomized Clinical Trial Evaluating AcceptME-A Digital Gamified Acceptance and Commitment Early Intervention Program for Individuals at High Risk for Eating Disorders.
Eating disorders (ED) constitute a serious public health issue affecting predominantly women and appearing typically in adolescence or early adulthood. EDs are extremely difficult to treat, as these disorders are ego-syntonic, and many patients do not seek treatment. It is vital to focus on the development of successful early-intervention programs for individuals presenting at risk and are on a trajectory towards developing EDs. This study is a randomized controlled trial evaluating an innovative digital gamified Acceptance and Commitment early-intervention program (AcceptME) for young females showing signs and symptoms of an ED and at high risk for an ED. Participants (n = 92; Mage = 15.30 years, SD = 2.15) received either AcceptME (n = 62) or a waitlist control (n = 30). Analyses indicated that the AcceptME program effectively reduced weight and shape concerns with large effects when compared to waitlist controls. Most participants scored below the at-risk cut-off (WCS score < 52) in the AcceptME at end-of-intervention (57.1%) compared to controls (7.1%), with odds of falling into the at-risk group being 14.5 times higher for participants in the control group. At follow-up, 72% of completers reported scores below the at-risk cut-off in the AcceptME group. The intervention also resulted in a decrease in ED symptomatology and increased body image flexibility. Overall, results suggest that the AcceptME program holds promise for early-intervention of young women at risk for developing an ED.


Cytochrome c oxidase binding of hydrogen peroxide.
Oxidized cytochrome c oxidase can bind hydrogen peroxide, as evidenced by changes in its spectrum and its ability to use hydrogen peroxide as an electron acceptor in cytochrome c oxidation. The affinity of the oxidized enzyme for hydrogen peroxide is high, with a Kd of less than 10 microM, and the binding is inhibited by ligands of cytochrome a3. Oxidized cytochrome c oxidase, in submitochondrial particles or solubilized in several ionic and nonionic detergents, binds peroxide with comparable affinities. The size of the spectral shift observed upon peroxide binding depends on the pH of the solution and differs in extinction coefficient between preparations, but all preparations tested appeared to bind peroxide. The differences in the magnitude of the spectral shift upon peroxide binding to different preparations suggest that oxidized cytochrome c oxidase as prepared may be made up of more than one species and that the proportion of the species which binds peroxide varies with the preparation. These studies of the binding of peroxide clarify the mechanism by which cytochrome c oxidase catalyzes the reduction of oxygen to water without the formation of free-radical intermediates.


SARS-CoV-2 mucosal vaccine protects against clinical disease with sex bias in efficacy.
Intranasal mucosal vaccines can more effectively induce mucosal immune responses against SARS-CoV-2. Here, we show in hamsters that an intranasal subunit mucosal vaccine boost with the beta variant S1 can prevent weight loss, in addition to reducing viral load, which cannot be studied in macaques that don't develop COVID-like disease. Protective efficacy against both viral load and weight loss correlated with serum antibody titers. A sex bias was detected in that immune responses and protection against viral load were greater in females than males. We also found that priming with S1 from the Wuhan strain elicited lower humoral immune responses against beta variant and led to less protection against beta viral challenge, suggesting the importance of matched antigens. The greater efficacy of mucosal vaccines in the upper respiratory tract and the need to consider sex differences in vaccine protection are important in the development of future improved COVID-19 vaccines.


Zuotai and HgS differ from HgCl2 and methyl mercury in Hg accumulation and toxicity in weanling and aged rats.
Mercury sulfides are used in Ayurvedic medicines, Tibetan medicines, and Chinese medicines for thousands of years and are still used today. Cinnabar (α-HgS) and metacinnabar (β-HgS) are different from mercury chloride (HgCl2) and methylmercury (MeHg) in their disposition and toxicity. Whether such scenario applies to weanling and aged animals is not known. To address this question, weanling (21d) and aged (450d) rats were orally given Zuotai (54% β-HgS, 30mg/kg), HgS (α-HgS, 30mg/kg), HgCl2 (34.6mg/kg), or MeHg (MeHgCl, 3.2mg/kg) for 7days. Accumulation of Hg in kidney and liver, and the toxicity-sensitive gene expressions were examined. Animal body weight gain was decreased by HgCl2 and to a lesser extent by MeHg, but unaltered after Zuotai and HgS. HgCl2 and MeHg produced dramatic tissue Hg accumulation, increased kidney (kim-1 and Ngal) and liver (Ho-1) injury-sensitive gene expressions, but such changes are absent or mild after Zuotai and HgS. Aged rats were more susceptible than weanling rats to Hg toxicity. To examine roles of transporters in Hg accumulation, transporter gene expressions were examined. The expression of renal uptake transporters Oat1, Oct2, and Oatp4c1 and hepatic Oatp2 was decreased, while the expression of renal efflux transporter Mrp2, Mrp4 and Mdr1b was increased following HgCl2 and MeHg, but unaffected by Zuotai and HgS. Thus, Zuotai and HgS differ from HgCl2 and MeHg in producing tissue Hg accumulation and toxicity, and aged rats are more susceptible than weanling rats. Transporter expression could be adaptive means to reduce tissue Hg burden.


Fatty acid composition of equine plasma.
Fatty acid composition of plasma lipids of normal horses was determined. Four fatty acids (C16:0, C18:0, C18:1, and C18:2) comprised 86.73% of the total, with C18:2 comprising 44.04% of the total. Eight other fatty acids were found in small amounts. Unsaturated fatty acids constituted 66% of the total. Marked variation was demonstrated in fatty acid occurrence and distribution in the sterol ester, triglyceride, phospholipid, and free fatty acid fractions.


A modern approach to olfactory groove meningiomas.
Management of olfactory groove meningiomas (OGMs) has changed significantly with the advances in extended endoscopic endonasal approaches (EEAs), which is an excellent approach for patients with anosmia since it allows early devascularization and minimizes retraction on the frontal lobes. Craniotomy is best suited for preservation of olfaction. However, not infrequently, a tumor presents after extending outside the reach of an EEA and a solely transcranial approach would require manipulation and retraction of the frontal lobes. These OGMs may best be treated by a staged EEA-craniotomy approach. In this study the authors' goal was to present their case series of patients with OGMs treated with their surgical approach algorithm. The authors conducted an IRB-approved, nonrandomized historic cohort including all consecutive cases of OGMs treated surgically between 2010 and 2020. Patient demographic information, presenting symptoms, operative details, and complications data were collected. Preoperative and postoperative tumor and T2/FLAIR intensity volumes were calculated using Visage Imaging software. Thirty-one patients with OGMs were treated (14 craniotomy only, 11 EEA only, and 6 staged). There was a significant difference in the distribution of patients presenting with anosmia and visual disturbance by approach. Tumor size was significantly correlated with preoperative vasogenic edema. Gross-total resection was achieved in 90% of cases, with near-total resection occurring twice with EEA and once with a staged approach. T2/FLAIR hyperintensity completely resolved in 90% of cases and rates did not differ by approach. Complication rates were not significantly different by approach and included 4 CSF leaks (p = 0.68). A staged approach for the management of large OGMs with associated anosmia and significant lateral extension is a safe and effective option for surgical management. Through utilization of the described algorithm, the authors achieved a high rate of GTR, and this strategy may be considered for large OGMs.


A novel role of RNA helicase A in regulation of translation of type I collagen mRNAs.
Type I collagen is composed of two α1(I) polypeptides and one α2(I) polypeptide and is the most abundant protein in the human body. Expression of type I collagen is primarily controlled at the level of mRNA stability and translation. Coordinated translation of α(I) and α2(I) mRNAs is necessary for efficient folding of the corresponding peptides into the collagen heterotrimer. In the 5' untranslated region (5' UTR), collagen mRNAs have a unique 5' stem-loop structure (5' SL). La ribonucleoprotein domain family member 6 (LARP6) is the protein that binds 5' SL with high affinity and specificity and coordinates their translation. Here we show that RNA helicase A (RHA) is tethered to the 5' SL of collagen mRNAs by interaction with the C-terminal domain of LARP6. In vivo, collagen mRNAs immunoprecipitate with RHA in an LARP6-dependent manner. Knockdown of RHA prevents formation of polysomes on collagen mRNAs and dramatically reduces synthesis of collagen protein, without affecting the level of the mRNAs. A reporter mRNA with collagen 5' SL is translated three times more efficiently in the presence of RHA than the same reporter without the 5' SL, indicating that the 5' SL is the cis-acting element conferring the regulation. During activation of quiescent cells into collagen-producing cells, expression of RHA is highly up-regulated. We postulate that RHA is recruited to the 5' UTR of collagen mRNAs by LARP6 to facilitate their translation. Thus, RHA has been discovered as a critical factor for synthesis of the most abundant protein in the human body.


Irradiation enhanced risks of hospitalised pneumonopathy in lung cancer patients: a population-based surgical cohort study.
Pulmonary radiotherapy has been reported to increase a risk of pneumonopathy, including pneumonitis and secondary pneumonia, however evidence from population-based studies is lacking. The present study intended to explore whether postoperative irradiation increases occurrence of severe pneumonopathy in lung cancer patients. The nationwide population-based study analysed the Taiwan National Health Insurance Research Database (covered >99% of Taiwanese) in a real-world setting. From 2000 to 2010, 4335 newly diagnosed lung cancer patients were allocated into two groups: surgery-RT (n=867) and surgery-alone (n=3468). With a ratio of 1:4, propensity score was used to match 11 baseline factors to balance groups. Irradiation was delivered to bronchial stump and mediastinum according to peer-audited guidelines. Hospitalised pneumonia/pneumonitis-free survival was the primary end point. Risk factors and hazard effects were secondary measures. Multivariable analysis identified five independent risk factors for hospitalised pneumonopathy: elderly (>65 years), male, irradiation, chronic obstructive pulmonary disease (COPD) and chronic kidney disease (CKD). Compared with surgery-alone, a higher risk of hospitalised pneumonopathy was found in surgery-RT patients (HR, 2.20; 95% CI, 1.93-2.51; 2-year hospitalised pneumonia/pneumonitis-free survival, 85.2% vs 69.0%; both p<0.0001), especially in elderly males with COPD and CKD (HR, 13.74; 95% CI, 6.61-28.53; p<0.0001). Unexpectedly, we observed a higher risk of hospitalised pneumonopathy in younger irradiated-CKD patients (HR, 13.07; 95% CI, 5.71-29.94; p<0.0001) than that of elderly irradiated-CKD patients (HR, 4.82; 95% CI, 2.88-8.08; p<0.0001). A high risk of hospitalised pneumonopathy is observed in irradiated patients, especially in elderly males with COPD and CKD. For these patients, close clinical surveillance and aggressive pneumonia/pneumonitis prevention should be considered. Further investigations are required to define underlying biological mechanisms, especially for younger CKD patients.