RNA sequencing (RNA-Seq) is becoming a popular method for measuring gene expression in non-model organisms, including wild populations sampled in the field. While RNA-Seq can be used to measure gene expression variation among wild-caught individuals and can yield important biological insights into organismal function, technical variables may also influence gene expression estimates. We examined the influence of multiple technical variables on estimated gene expression in a non-model fish species, the westslope cutthroat trout (Oncorhynchus clarkii lewisi), using two RNA-Seq methods: 3’ RNA-Seq and whole mRNA-Seq. We evaluated the effects of dip netting versus electrofishing, and of harvesting tissue immediately versus 5 minutes after euthanasia on estimated gene expression in blood, gill, muscle, and liver. We found higher RNA degradation in the liver compared to the other tissues. There were fewer expressed genes in blood compared to gill and muscle. We found no difference in gene expression among sampling methods or due to a delay in tissue collection. However, we detected fewer genes with 3’ RNA-Seq than with whole mRNA-Seq and found statistically significant differences in gene expression between 3’ RNA-Seq and whole mRNA-Seq. The magnitude and direction of these differences does not appear to be dependent on gene type or length. Our findings indicate that RNA-Seq is robust to the technical variables related to the field sampling techniques tested here but varies based on the tissue sampled and the RNA-Seq library used. This study advances understanding of usefulness of RNA-Seq to study gene expression variation in evolution, ecology, and conservation.

About 30% of patients with epilepsy do not respond to anti-epileptic drugs leading to refractory seizures. The pathogenesis of drug-resistance in Mesial Temporal Lobe Epilepsy (MTLE) is not completely understood. Increased activity of drug-efflux transporters might be involved, resulting in subclinical concentrations of the drug at the target site. The major drug-efflux transporters are permeability glycoprotein (P-gp) and multidrug-resistance protein-1 (MRP-1). We have studied these two transporters in the sclerotic hippocampal tissues resected from the epilepsy surgery and compared their expression profile with the tissues resected from non-epileptic autopsy cases. Statistically significant over expression of both P-gp (p-value<0.0001) and MRP-1 (p-value 0.01) at gene and protein levels was found in the MTLE cases. The fold change of P-gp was more pronounced than MRP-1. Immunohistochemistry of patient group showed increased immunoreactivity of P-gp at blood brain barrier and increased reactivity of MRP-1 in parenchyma. The results were confirmed by confocal immunofluorescence microscopy. This suggested that P-gp in association with MRP-1 might be responsible for the multi-drug resistance in epilepsy.

Maternal diabetes insipidus does not usually adversely affect the course of pregnancy. We present a rare case of central diabetes insipidus diagnosed at 31 weeks of gestation with fetal oligohydramnios successfully treated with intranasal desmopressin. To date, only three cases of diabetes insipidus with oligohydramnios have been reported.

Aim To undertake a scoping review of pharmacists’ activities in opioid medicines management in primary care settings, including those developed or led by pharmacists, or in which pharmacists were members of broader multi-disciplinary teams; and to collate the activities, models of care and settings, and reported outcomes. Methods The bibliographic databases MEDLINE, EMBASE, International Pharmaceutical Abstracts, CINAHL, SCOPUS and Web of Science were searched. Studies with quantitative evaluation and published in English were eligible. Participants were patients with any pain category or an opioid use disorder, and healthcare providers. Studies originating in hospitals or involving supply functions were not included. Screening of literature and data charting of results were undertaken by two researchers. Results The 47 studies included in the scooping review occurred in primary care settings collated into four categories: general practice or primary care clinics; healthcare organisations; community pharmacies and outreach services. Studies were primarily of opioid use in chronic, non-cancer pain. Other indications were opioid use disorder, cancer and dental pain. Pharmacist activities targeted risk-mitigation, patient and provider education and broader, strategic approaches. Patient-related outcomes included reduced opioid load, improved functionality and symptom management, enhanced access to services and medication-assisted treatments, and engagement in risk-mitigation strategies. Behaviour change of providers was demonstrated. Conclusion The review has identified the significant contribution that pharmacists working in primary care settings can make to minimise harm from opioids. Strategies implemented in isolation have the potential to further reduce adverse clinical outcomes with greater collaboration and coordination, such as opioid stewardship.

Global change drivers like warming and changing nutrient cycles have a substantial impact on ecosystem functioning. In most modelling studies, organism responses to warming are described through the temperature dependence of their biological rates. In nature, however, organisms are more than their biological rates. Plants are flexible in their elemental composition (stoichiometry) and respond to variance in nutrient availability and temperature. An increase in plant carbon-to-nutrient content means a decrease in food quality for herbivores. Herbivores can react to this decrease by compensatory feeding, which implies higher feeding rates and higher carbon excretion to optimize nutrient acquisition. In a novel model of a nutrient-plant-herbivore system, we explored the consequences of flexible stoichiometry and compensatory feeding for plant and herbivore biomass production and survival across gradients in temperature and nutrient availability. We found that flexible stoichiometry increases plant and herbivore biomasses, which results from increased food availability due to higher plant growth. Surprisingly, compensatory feeding decreased plant and herbivore biomasses as overfeeding by the herbivore reduced plants to low densities and depleted their resource. Across a temperature gradient, compensatory feeding caused herbivore extinction at a lower temperature, while flexible stoichiometry increased its extinction threshold. Our results suggest that compensatory feeding can become critical under warm conditions. In contrast, flexible stoichiometry is beneficial for plants up to a certain temperature threshold. These findings demonstrate the importance of accounting for adaptive and behavioural organismal responses to nutrient and temperature gradients when predicting the consequences of warming and eutrophication for population dynamics and survival.

Despite the potential of tissue engineering approaches for cartilage repair, a major shortcoming is the low biosynthetic response of chondrocytes. While different strategies have been investigated to upregulate tissue formation, a novel approach may be to control nutrient metabolism. Although known for their anaerobic metabolism of glucose, chondrocytes are more synthetically active when cultured under conditions that elicit mixed aerobic-anaerobic metabolism. Here, we postulate this metabolic switch induces hypoxia inducible factor 1α (HIF-1α) signaling leading to improved tissue growth. Transition to different metabolic states can result in the pooling of intracellular metabolites, several of which can stabilize HIF-1α by interfering with proline-hydroxylase-2 (PHD2). Chondrocytes cultured under increased media availability accelerated tissue deposition (2.2 to 3.5-fold) with the greatest effect occurring at intermediate volumes (2 mL/106 cells). Under higher media volumes, metabolism switched from anaerobic to mixed aerobic-anaerobic. At and beyond this transition, maximal changes in PHD2 activity (- 45%), HIF-1α protein expression (8-fold increase), and HIF-1 gene target expression were observed (2.0 to 2.7-fold increase). Loss-of-function studies using YC-1 (to degrade HIF-1α) confirmed the involvement of HIF-1 signaling under these conditions. Lastly, targeted metabolomic studies of glucose metabolites (14 in total) revealed that both intracellular lactate and succinate correlated with PHD2 activity. Although both metabolites can inhibit PHD2, this effect can most likely be attributed to lactate as succinate was only present in trace amounts. However, addition work (e.g., 13C flux analyses) are required to confirm this assertion. Nevertheless, by harnessing this newly identified metabolic switch, functional engineered cartilage implants may be developed without the need for sophisticated methods which would allow for improved translation into the clinical realm.

In this paper, we study the following kind of Schr\”{o}dinger-Poisson system in ${\R}^{2}$ \begin{equation*} \left\{\begin{array}{ll} -\Delta u+V(x)u+\phi u=K(x)f(u),\ \ \ x\in{\R}^{2},\\ -\Delta \phi=u^{2},\ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ x\in{\R}^{2}, \end{array}\right. \end{equation*} where $f\in C({\R}, {\R} )$, $V(x)$ and $K(x)$ are both axially symmetric functions. By constructing a new variational framework and using some new analytic techniques, we obtain an axially symmetric solution for the above planar system. our result improves and extends the existing works.

In this paper, the anti-plane shear motion of an asymmetric three-layered inhomogeneous elastic plate has been examined. An asymptotic approach is employed for the present investigation. Both the generalized and unified dispersion relations within the long-wave low-frequency range have been determined. The obtained unified dispersion relation is investigated taking into account the recently analyzed material contrast for layered plate with mixed stiff-soft layers of different material properties. Finally, we make comparison with symmetric plate being a special case of the asymmetric plate under consideration in the end.

Soil properties play an important role in rill development and erosion. In this investigation, rill morphology developmental processes under sandy loam (SL), light loam (LL), medium loam (ML) and heavy loam (HL) soils on the Loess Plateau, China, were compared using laboratory experiments. Experimental analysis included two rainfall intensities (90 and 120 mm/h) and four slope treatments (0°, 15°, 20° and 25%). Results indicate that HL is the most prone to rill development, and SL, LL and ML are prone to rill development under heavy rain, with SL rill erosion being the most sensitive to heavy rain. The development of rills in SL are mainly characterized by an increase in rill width and merging nodes; rills in HL were mainly characterized by an increase in rill length, merging nodes and rill number. LL and ML rill development indices were between SL and HL. Differences in runoff collection caused by rill morphology differences further promoted differences in soil erosion. Rainfall intensity has a positive effect on rill shape parameters of all soils; slope has a positive and negative double effect on SL, LL and ML rill shape parameters, and only a positive effect on HL rill shape parameters. The sensitivity of rill parameters to rainfall intensity and slope angle depends on soil infiltration performance, surface soil stability and soil structure stability. Based on soil characteristic factors and rill morphological parameters, an empirical model of slope erosion in the loess region was established.

Rituximab use in multiple sclerosis has been promising. Cytokine release syndrome (CRS) is a common side effect of rituximab in patients with lymphoma. We report a case of a 44-year-old man with a history of relapsing-remitting multiple sclerosis, who presented with signs and symptoms consistent with CRS after rituximab initiation.

The time-global unique solvability on the reaction--diffusion equations for preypredator models and dormancy on predators is established. The crucial step is to construct time-local nonnegative classical solutions by using a new approximation associated with time-evolution operators. Although the system does not equip usual comparison principles, a priori bounds are derived, so solutions are extended time-globally. Via observations to the corresponding ordinary differential equations, invariant regions and asymptotic behaviors of solutions are also investigated.

In silico search for planar hexacoordinate silicon center has been initiated by global minimum screening with density functional theory and energy refinement using coupled cluster theory. The search resulted in a local minimum of SiAl3Mg3H2+ structure which contains a planar hexacoordinate silicon center (phSi). The phSi structure is 5.8 kcal/mol higher in energy than the global minimum. However, kinetic studies reveal that the local minimum structure has enough stability to be detected experimentally. Born-Oppenheimer molecular dynamics (BOMD) simulations reveal that the phSi structure can be maintained up to 400 K. The formation of multiple bonds between the central silicon atom and framework aluminium atom is the key stabilizing factor for the planar structure.

An 83 year old female had asymptomatic SARS-CoV-2 infection while taking ruxolitinib. She remained RT-PCR positive for viral RNA for >120 days, Pegylated interferon for 4 weeks led to viral RNA clearance. The observations support combination therapy of ruxolitinib + interferon for COVID-19.

In this article we show the existence, uniqueness and Ulam stability results of the solution for a class of a nonlinear Caputo fractional integro-differential problem with mixed conditions. we use three fixed point theorems to proof the existence and uniqueness results. By the results obtained, the reasons for the Ulam stability are verified. An example proposed to illustrate our main results.

Germline mutations are the raw material for natural selection, driving species evolution and the creation of earth’s biodiversity. Life on earth would stagnate without this driver of genetic diversity. Yet, it is a double-edged sword. An excess of mutations can have devastating effects on fitness and population viability. It is therefore one of the great challenges of molecular ecology to determine the rate and spectrum by which these mutations accrue across the tree of life. Advances in high-throughput sequencing are providing new opportunities for characterizing these rates and patterns within species and populations, thus informing essential evolutionary parameters such as the timing of speciation events, the intricacies of historical demography, and the degree to which lineages are subject to the burdens of mutational load. Here, we will focus on the applications and limitations of whole-genome comparisons among closely related individuals in what are typically described as “trio” analyses for the detection of germline mutations as they arise in real time. By sequencing and comparing whole-genomes generated for individuals of known relatedness – typically, parent to offspring – investigators can ideally count and characterize mutations as they appear per generation. The promise for gaining insight into classic hypotheses of molecular evolution is high, though so too is the cost. Namely, the technical challenges are daunting given that pedigree-based studies are essentially searching for needles in a haystack. Even so, the opportunities are so enticing, and the field so young, we can say with confidence that fundamental insights have only just begun to emerge.

Carotenoid-based colours are a textbook example of honest signalling because carotenoids must be acquired from the environment. However, many species produce similar colours using self-synthesised pteridine pigments. A compelling but untested hypothesis is that pteridines compensate for low environmental availability of carotenoids because it is metabolically cheaper to synthesise pteridines than to acquire and sequester carotenoids. Based on a phylogenetic comparative analysis of 11 pigment concentrations in skin tissue of agamid lizards, we show that pteridine concentrations are higher and carotenoid concentrations lower in less productive environments. Both carotenoid and pteridine pigments were present in all species, but only pteridine concentrations explained colour variation among species. Furthermore, pigment concentrations were uncorrelated with indices of sexual selection. These results suggest that variation among species in pteridine synthesis compensates for environmental availability of carotenoids and challenge the paradigm of honest carotenoid signalling in vertebrates with complex colour production mechanisms.

Commentary:

Background and Purpose: Individualized assessment of the activity of cytochrome P450 2D6 (CYP2D6), a highly variable drug-metabolizing enzyme, is performed through phenotyping during which a probe drug is administered to measure the enzyme’s activity. In order to avoid any iatrogenic harm (allergic drug reaction, dosing error) related to the probe drug, the development of non-invasive tools for real-time phenotyping of CYP2D6 could significantly contribute to the expansion of precision medicine in clinical practice. This study focuses on the identification of endogenous markers of the CYP2D6 enzyme in human biofluids using a liquid chromatography (LC)-high-resolution mass spectrometry (HRMS)-based metabolomics approach. Experimental Approach: Data from a control session were compared to data from an inhibition session. Before the latter, healthy volunteers (extensive and ultrarapid metabolizers) received a daily dose of paroxetine 20 mg over seven days. CYP2D6 genotyping and phenotyping, using single oral dose of dextromethorphan 5 mg, were also performed in all participants. Key Results: In CYP2D6 extensive and ultrarapid metabolizers (n = 37), mean relative intensities of five features were significantly reduced during the inhibition session compared to the control session (fold changes ≤ 0.67, FDR-adjusted P < 0.0001). Furthermore, mean relative intensities of these candidates were significantly higher in the CYP2D6 extensive-ultrarapid metabolizer group (n = 37) compared to the poor metabolizer group (n = 6) (fold changes ≤ 0.67, P < 0.0001). Conclusion and Implications: The applied untargeted metabolomics strategy was able to identify five CYP2D6 endogenous metabolites, a promising discovery for non-invasive phenotyping and personalised medicine.

The pandemic caused by the novel coronavirus SARS-CoV2 has led to significant morbidity and mortality. Susceptibility to either contracting infection or presenting with symptoms has varied to a great extent among different populations. Whether specific occupations or lifestyles, other than the ones that predispose individuals to increased exposure, also play a role in shaping infection susceptibility is not yet fully understood. In India, due to a rich heritage of different classical dance genres, dance vocationally engages a great number of people. Moreover dance, being an aesthetic rendition of body movements, is established to have major influence of human health. With an aim to assess the susceptibility of SARS-CoV2 infection among the practitioner of various Indian classical dance genres, an internet-based survey was done based on a questionnaire. 6.04% of the dancers participating in the survey (N=182) reported past infection with SARS-CoV-2 virus confirmed by RT-PCR test, while 15.38% of them reported the same for one or more household members. The survey did not find any significant difference in age, body mass index between infected versus non-infected individuals. Further exploration of this occupational group with respect to COVID-19 disease outcomes as well as lung functions in health and disease are warranted.

Introduction Residual stone fragments remain a challenging topic for urologists following retrograde intrarenal surgery (RIRS). This study investigates the effectiveness of second-look flexible ureterorenoscopy (URS) to achieve a true stone-free status and decrease stone-related events. Material and Method A total of 176 consecutive patients treated with RIRS for kidney stones were included in the study. Patients were divided into two groups, group 1 receiving the standard of care (n=51) and group 2 receiving second-look flexible URS (n=125). In all cases, one or more calyceal stones of 1 to 4 mm were extracted with a basket at the time of stent removal. Unless earlier intervention was necessary, patients were followed up every six months to determine if they had experienced SREs. Results The mean follow-up time for the whole group was 21±11.1 months. The mean number of stones in group 1 and group 2 was 1.25±0.11 and 1.56±0.19, respectively. Postoperative stone-free rate after RIRS for group 1 and group 2 were 37.25% (n=19/51) and 40.8% (n=51/125), respectively. The SFR improved to 93.6% (n=117/125) in group 2. Multivariable analysis showed that type of intervention, stone size, and body mass index were independent prognostic factors for SREs. When group 2 was taken as a reference, the odds ratio for SREs was 8.48 (95% CI: 2.95–24.42) in group 1. Conclusion This study found that second-look flexible URS increased the SFRs and diminished the number of SREs. We propose performing second-look flexible URS following RIRS in the presence or suspicion of RSFs to provide better treatment results, less radiation exposure, and greater patient satisfaction. Key Words Retrograde Intrarenal Surgery, Flexible Ureteroscopy, Stone-free Rate, Residual stone Fragments, Stone-Related Event, Second-look Flexible Ureteroscopy.

Objective: To present an alternative surgical technique in treating cases of Chiari I Malformation with mild-to-moderate syringomyelia after decompressive suboccipital craniectomy: incising only the outer layer of the dura mater, then dissecting it from the inner layer without opening the latter. Methods and Results: We utilized this technique in a short series of three cases who were admitted in our department for mild symptoms such as intermittent headache and dissociated sensory loss in the upper limbs, caused by a Chiari Malformation Type I. The patients were placed in the sitting position. We performed a reduced median suboccipital craniectomy and resection of the posterior arch of C1 adapted to the level of tonsil descent, from a limited superior half to a complete resection. Afterwards, we incised the outer dural layer, while sparing the inner one. Using a fine dissector, we then split apart the outer and inner layers to the margin of the craniectomy. Through the transparency of the inner layer and the arachnoid, the cerebellum and the medulla were visible and pulsating. An autologous fascia duraplasty was then performed. The postoperative course was favorable in all cases, patients being discharged without any deficits and with complete symptom resolution. Conclusions: Interlayer dural split technique can be used effectively in treating symptomatic cases of type I Chiari malformation in adults, with mild-to-moderate syringomyelia. It is less invasive than opening the dura and possibly more effective than decompressive craniectomy and C1 laminectomy alone. This technique must be validated in a larger case-control series.

Ischemic mitral regurgitation (IMR) is a common complication, which is accompanied by myocardial infarction, causing heart failure and leading to poor prognosis. Although several surgical techniques have been reported, certain surgical methods have not been established for treating IMR. We report a successful case of left ventricular posterior wall plication through a left atriotomy over the mitral valve for IMR in a patient who experienced cardiac shock and could not be weaned off mechanical support. Posterior wall plication changed the left ventricle from a spherical to an oval shape, restored the position of papillary muscles and posterior wall, improved leaflet tethering, and prevented further remodeling of the left ventricle. This method may be useful for treating IMR and improve patients’ prognosis.

Background: Muscle wasting and limitation in physical activity have been reported in patients with hemodialysis; this leads to poor quality of life at the end. Objective: To develop the respiratory training device and to determine the efficiency of the prototype on respiratory muscle strength, functional capacity and dyspnea perception in hemodialysis patients. Design, setting and participants: the development of respiratory device was created and then effects of respiratory device were examined in hemodialysis patients. A total of 25 patients with quasi experimental study was recruited in the present study with aged ≥ 35 years old. All participants had a history of hemodialysis 3 days per week. Outcome measurements and statistical analysis: The protocol was conducted 15 inhales, three set per day with 40% of maximal inspiratory pressure; totally 45 inhales. The participants were asked to perform for three times per week during on hemodialysis. Respiratory muscle strength, the 6minute walk test and rate of dyspnea scores were assessed before and after 8-week intervention program. Paired t-tests was used to compare between initial values and follow-up values in respiratory prototype. Results: Of 25 patients at initially was enrolled, 22 individuals (88%) completed in the 8-week program. Significant improve in inspiratory muscle strength, 6minite walk distance and rate of dyspnea was observed after 8-week intervention (12.44± 3.55 cmH2O, 24.78 ± 8.89 meters, and 0.50 ± 0.19, respectively). Conclusion: Using porotype of respiratory device can effectively improve cardiorespiratory performance which is marked by increasing inspiratory muscle functional capacity and rate of dyspnea in hemodialysis patients after 8-week training program.

Objective: This study aimed to investigate the association between carotid intima-media changes that play a part in the atherosclerotic process in childhood obesity and fibrin monomers as an important indicator of fibrin plaque. Methods: This is a cross-sectional study of obese children and non-obese healthy control subjects. Height, weight, body mass index, waist/hip ratio, systolic/diastolic blood pressures were recorded, in addition, biochemistry, hemogram, fibrin monomers and d-dimer were measured in both groups. Right and left common carotid intima-media thicknesses were measured by ultrasonography and mean carotid intima-media thickness was calculated. Results: Obese children (n=89, 46.1% girls, median age: 12.6±2.3 years) and healthy control group (n=40, 52.5% girls, median age: 13.2±2.2 years) were comparable in terms of gender, age and puberty stage. Mean carotid intima-media thickness was higher in obese children than the healthy control group (p=0.002). There was no difference between the two groups in terms of fibrin monomers and D-dimer levels. In obese children, there was a weak negative correlation between mean carotid intima-media thickness and fibrin monomers (p=0.030, r=-0.233). Conclusion: In obese children, mean carotid intima-media thickness was determined higher, as an early indicator of atherosclerosis. We want to emphasize that obese children are at risk for cardiovascular disease and should be evaluated in terms of atherosclerosis. This study investigates the relation between increased carotid intima-media thickness and fibrin monomers, in children, the first time in Literature. What’s already known about this topic? It is possible to reveal the early period of the atherosclerosis process by showing carotid intima medial thickness. Fibrin is a major component of many atherosclerotic plaques. What does this article add? Our study investigated the relationship between mean carotid intima-media thickness in childhood obesity and fibrin monomers. But no positive correlation was found between fibrin monomers and the carotid intima-media thickness.