Combination of docking, molecular dynamics and quantum mechanical calculations for metabolism prediction of 3,4-methylenedioxybenzoyl-2-thienylhydrazone.
In modern drug discovery process, ADME/Tox properties should be determined as early as possible in the test cascade to allow a timely assessment of their property profiles. To help medicinal chemists in designing new compounds with improved pharmacokinetics, the knowledge of the soft spot position or the site of metabolism (SOM) is needed. In silico methods based on docking, molecular dynamics and quantum chemical calculations can bring us closer to understand drug metabolism and predict drug-drug interactions. We report herein on a combined methodology to explore the site of metabolism prediction of a new cardioactive drug prototype, LASSBio-294 (1), using MetaPrint2D to predict the most likely metabolites, combined with structure-based tools using docking, molecular dynamics and quantum mechanical calculations to predict the binding of the substrate to CYP2C9 enzyme, to estimate the binding free energy and to study the energy profiles for the oxidation of (1). Additionally, the computational study was correlated with a metabolic fingerprint profiling using LC-MS analysis. The results obtained using the computational methods gave valuable information about the probable metabolites of (1) (qualitatively) and also about the important interactions of this lead compound with the amino acid residues of the active site of CYP2C9. Moreover, using a combination of different levels of theory sheds light on the understanding of (1) metabolism by CYP2C9 and its mechanisms. The metabolic fingerprint profiling of (1) has shown that the metabolites founded in highest concentration in different species were metabolites M1, M2 and M3, whereas M8 was found to be a minor metabolite. Therefore, our computational study allowed a qualitative prediction for the metabolism of (1). The approach presented here has afforded new opportunities to improve metabolite identification strategies, mediated by not only CYP2C9 but also other CYP450 family enzymes.


Glutamate-induced cell death in HT22 mouse hippocampal cells is attenuated by paxilline, a BK channel inhibitor.
In the present study, we show that the large conductance calcium-activated potassium channel (BK(Ca) channel) inhibitor paxilline protects neuronal cells against glutamate-induced cell death. In our studies, we used HT22 mouse hippocampal cells as an experimental model and observed that the effect of paxilline was dose-dependent. We also found that other inhibitors of BK(Ca) channels, iberiotoxin and charybdotoxin, were not cytoprotective. Paxillinol, which is a structural analog of paxilline but does not inhibit BK(Ca) channel, also protected HT22 cells against glutamate-induced toxicity. These data suggest that the observed cytoprotection was not related to BK(Ca) channel inhibition by paxilline. In addition, paxilline neither restored glutathione levels nor reduced the amount of reactive oxygen species upon glutamate treatment. Our results suggest that paxilline protects neuronal HT22 cells against glutamate-induced cell death independently of BK(Ca) channel activity and oxidative stress induced by glutamate treatment.


COVID-19: leadership on the frontline is what matters when we support healthcare workers.
The implications of severe acute respiratory syndrome coronavirus (SARS-CoV-2, henceforth described as COVID-19) on healthcare systems globally are proving to be immense, with unforeseen impacts that are still to fully emerge. Local and national healthcare systems, hospitals and healthcare workers have been overwhelmed by the needs of patients and limited by weaknesses in resources, staff capacity and distribution networks. These circumstances impact the ergonomic conditions within which healthcare staff work and subsequently their behavioural responses.In this commentary, we argue that urgent research is needed globally to bridge the evidence gap that exists on how best to support healthcare workers with the repercussions of working on the frontline of a pandemic. Leadership on the frontline is what matters. It is not only what policies, guidelines and checklists are in place to support nurses, doctors and healthcare workers, it is the actions and behaviours of their frontline and local leaders in implementing initiatives that really make the difference.Recognizing that the leadership style, organizational culture and model of successful implementation are inextricable is the first step to ensure sustainable interventions to support healthcare workers' well-being will follow.


[Retention and biomechanics of retentive complexes. 3. The Kratochvil school and current retentive complexes].
The authors analyse the biomechanical bases of the Kratochvil "retentive complex" and show how they have induced the nowadays north-american propositions (RPI & RPA). They compare them to the european ones (Nally-Martinet).


Trophic interactions and habitat segregation between competing Daphnia species.
Two commonly coexisting species of Daphnia segregate by habitat in many stratified lakes. Daphnia pulicaria is mostly found in the hypolimnion whereas D. galeata mendotae undergoes diel vertical migration between the hypolimnion and the epilimnion. I examined how habitat segregation between these two potentially competing species might be affected by trophic interactions with their resources and predators by performing a field experiment in deep enclosures in which I manipulated fish predation, nutrient levels, and the density of epilimnetic Daphnia. The results of the experiment indicate that habitat use by D. pulicaria can be jointly regulated by competition for food from epilimnetic Daphnia and predation by fishes. Patterns of habitat segregation between the two Daphnia species were determined by predation by fish but not by nutrient levels: The removal of epilimnetic fish predators resulted in higher zooplankton and lower epilimnetic phytoplankton densities and allowed D. pulicaria to expand its habitat distribution into the epilimnion. In contrast, increased resource productivity resulted in higher densities of both Daphnia species but did not affect phytoplankton levels or habitat use by Daphnia. The two species exhibit a trade-off in their ability to exploit resources and their susceptibility to predation by fish. D. g. mendotae (the less susceptible species) may thus restrict D. pulicaria (the better resource exploiter) from the epilimnion when fish are common due to lower minimum resource requirements than those needed by D. pulicaria to offset the higher mortality rate imposed by selective epilimnetic fish predators. D. g. mendotae does not appear to have this effect in the absence of fish.


Prediction, Characterization, and In Silico Validation of Chimeric RNAs.
Many chimeric RNA prediction software packages are available to assist the scientific community in searching for cancer-specific chimeric RNAs. These packages predict a large number of false positive events, which significantly hampers experimental validation of predicted chimeric RNAs. Here, we describe the detailed steps for (1) prediction of chimeric RNAs using EricScript software, (2) characterization of chimeric RNAs to discard most probable false positive events, and (3) in silico validation of chimeric RNA to select the potential cancer-specific events.


Realgar‑induced KRAS mutation lung cancer cell death via KRAS/Raf/MAPK mediates ferroptosis.
KRAS is a biomarker for non‑small cell lung cancer‑targeted therapy, but there is currently no effective KRAS‑targeting medication. Realgar is an impelling anticancer drug, however its significance in KRAS mutant lung cancer is uncertain. According to our findings, the IC50 of H23 (KRAS mutant) cells is 2.99 times lower than that of H1650 (non‑KRAS mutant) cells. Flow cytometry and the Hoechst 33258 staining assay revealed that H1650 cells treated with 4 µg/ml realgar had an apoptotic rate of 8.2%, while H23 cells had a rate of 21.46%. Accordingly, realgar was more sensitive to KRAS mutant cells. Transcriptome sequencing test indicated that there were 481 different expression genes in H23 cells treated with realgar. In H23 cells treated with realgar, mitochondria shrank, inner membrane folding was disturbed, and mitochondrial membrane potential crushed. Realgar boosted intracellular Fe2+, reactive oxygen species, malondialdehyde and glutathione levels, which were all reversed by ferroptosis inhibitor Fer‑1. Realgar decreased phosphorylated p‑Raf, p‑ERK1/2 and increased p‑p38 and p‑JNK, whereas only p‑Raf was abolished by Fer‑1. Raf inhibitor Sorafenib accelerated the realgar‑induced ferroptosis. On H23 cells treated with realgar, the expression of GPX4, SCL7A11 decreased while ACSL4 expression increased; this effect could also be amplified by Sorafenib. In conclusion, the present study indicated that realgar may induce ferroptosis by regulating the Raf, and hence plays a role in anti‑KRAS mutant lung cancer.


Cognitively engaging movement games improve interference control and academic performance in overweight children: A randomized control trial.
The purpose of this study was to determine the (a) dose-response effects of cognitively engaging movement games (CEMGs) designed to improve aerobic capacity, interference control (IC), and academic performance, (b) relationship between intervention-induced improvements in aerobic capacity, IC, and academic performance, and (c) moderation effect of IC on the relationship between aerobic capacity and academic performance in overweight children. Seventy-five overweight children (aged 11.23 ± 0.60 years; 48% males) participated in this study conducted in Taipei during the 2018/2019 academic year and were randomly assigned to the low-dose (20-min) intervention, high-dose (40-min) intervention, and control groups and completed a Stroop test, half-mile run, and language and mathematics tests before and after a 10-week afterschool program. Both intervention groups showed similar improvements in outcomes of aerobic capacity (ds > 0.80), IC (ds > 0.76), and academic performance (ds > 0.90) from the pretest to posttest, whereas these outcomes were unchanged for the control group. Furthermore, improved IC moderated the association between improvements in aerobic capacity and academic performance. Our findings suggest that CEMG with varying doses is feasible and effective for improving aerobic capacity, IC, and academic performance in overweight children and that the association between improvements in aerobic capacity and academic performance depends on the intervention effects on IC.


Artificial Intelligence for Oncology Nursing Authors: Potential Utility and Concerns About Large Language Model Chatbots.
Artificial intelligence is a revolution in the computing and data scientist era that has led to excitement and controversy in many fields, including research and publishing. As we move further into the artificial intelligence.


Cerebrospinal Fluid Lactate and Glucose Levels as Predictors of Symptomatic Delayed Cerebral Ischemia in Patients with Aneurysmal Subarachnoid Hemorrhage.
Aneurysmal subarachnoid hemorrhage (aSAH) is a complex neurovascular syndrome with profound systemic effects associated with high rates of disability and mortality. Delayed cerebral ischemia (DCI), which encompasses all neurobiological events occurring in the subacute-late stage after aSAH, has a complex pathogenesis and can occur in the absence of instrumental vasospasm. Our aim was to assess the correlation between cerebrospinal fluid (CSF) lactate and glucose levels measured on the second or third day after aSAH with clinical deterioration caused by DCI and with 3-month functional outcome. This prospective study included all aSAH patients admitted between January 2020 and December 2021 who underwent external ventricular drain placement and CSF lactate and glucose measurement. Among 133 aSAH patients, 48 had an external ventricular drain placed and early CSF lactate and glucose assessment. Independent predictors of symptomatic DCI were World Federation of Neurosurgical Societies grade IV-V (adjusted odds ratio [aOR] 25.8, 95% confidence interval [CI] 2.9-649.2, P = 0.012), elevated CSF glucose (aOR 28.8, 95% CI 3.3-775.2, P = 0.010), and elevated CSF lactate (aOR 14.7, 95% CI 1.9-205.7, P = 0.018). The only independent predictor of 3-month functional outcome was occurrence of symptomatic DCI (aOR 0.02, 95% CI 0.0-0.2, P = 0.01). Elevated CSF lactate and glucose levels in the first 3 days following aSAH were independent predictors of subsequent DCI-related neurological impairment; the presence of instrumental vasospasm was not significantly correlated with DCI after multivariate adjustment. CSF lactate and glucose monitoring may represent a point-of-care test, which could potentially improve prediction of subacute neurological worsening and guide therapeutic choices. Further research with larger prospective cohorts is warranted.


Solid emulsion gel as a novel construct for topical applications: synthesis, morphology and mechanical properties.
A series of the solid emulsion gels with the oil volume fraction in the range of 0-50% were synthesized through a polycondensation reaction between activated p-nitrophenyl carbonate poly(ethylene glycol) and protein-stabilized oil-in-water emulsions. The resultant structures were investigated in terms of swelling behavior, composition, morphology, mechanical and skin hydration properties. Solid emulsions gels share the properties of both hydrogel and emulsion. Similar to the classical hydrogel, the SEG swells in water up to equilibrium swelling degree, which decreases as the oil volume fraction increases, and comprises immobilized drops of protein-stabilized oil. The impregnation of the oil phase is found to reduce tensile stiffness of the material, but improves material's extensibility. The mechanical properties of the constructs (Young moduli in the range of 9-15 kPa and the elongation at break of 120-220%) are interpreted according to the "rule of elasticity mixture" that considers the elasticity of the composite material to be a sum of the contributions from individual components, i.e. hydrogel and dispersed oil drops. An idealized model that takes into account the history of the material preparation has been proposed to explain the improved extensibility of the constructs. The results of the mechanical tests, equilibrium swelling, and the skin hydration effect of the solid emulsion gels in vivo are discussed from the perspective of the biomedical applications of the solid emulsion gels, in particular, for the transdermal delivery of hydrophilic and lipophilic drugs.


Seasonal changes in circulating levels of luteinizing hormone and growth hormone in the migratory Canada goose.
Seasonal changes in plasma levels of LH and GH in the Canada goose (Branta canadensis interior) have been studied. The population examined breeds in the Canadian subarctic region (Fort Churchill, Manitoba) and winters in the southern United States (Swan Lake Refuge, Missouri). Peak levels of LH were observed during spring postmigratory (nestbuilding, early May) and nesting (incubating, early June) periods, while no significant differences were observed during the other periods studied. The highest levels of GH were observed during molting (late July) and the immediately following fall premigratory (early September) period.


Effect of glutamate and angiotensin II on whole cell currents and release of nitric oxide in the rat subfornical organ.
Blood-borne angiotensin II (AngII) is known to mediate water-intake by its excitatory effect on neurons in the subfornical organ (SFO). Conversely, nitric oxide (NO) has exclusively inhibitory effects on rat SFO-neurons and on SFO-mediated water-intake. Extracellular and patch-clamp recordings from freshly dissociated rat SFO-neurons showed that glutamate activates AngII-sensitive SFO-neurons by opening ligand-gated cation channels. An immunocytochemical study showed that activation of glutamate receptors increased the concentration of the inhibitory second messenger cGMP in the SFO. A model is proposed suggesting that NO protects SFO-neurons from overexcitability by excitatory neurotransmitters.


Protein impurities from cell culture dramatically impact transduction efficiency of polymer/virus hybrid vectors.
Polyethylenimine (PEI) was used recently with murine leukemia virus-like particles (MLV-VLPs) to produce a hybrid vector that possesses advantages over the native virus; the transduction efficiency of this vector, however, was less than the transduction efficiency of the native virus. The cause of the reduced efficiency was hypothesized to be related to the involvement of proteins in PEI/MLV-VLP complex formation and overall complex size. To test the hypothesis and potentially improve the efficiency of the hybrid vector, ultracentrifugation and size exclusion chromatography were used to purify MLV-VLP and to study the effect of proteins in cell culture medium on complex formation. Based on dynamic light scattering and electron microscopy, complexes formed from the purified MLV-VLPs were smaller, but surprisingly, less efficient than complexes formed from unpurified MLV-VLPs. The addition of protein to purified MLV-VLPs showed that the initial efficiency could be restored and that the purification strategy was not inactivating the MLV-VLPs. Further, by optimizing the amount of protein added to the purified MLV-VLPs, the level of transduction by PEI/MLV-VLP improved 1.6-fold. Particle characterization showed a correlation between the size of the PEI/MLV-VLP complex and the transduction efficiency, which is likely a result of greater sedimentation and cell contact during in vitro studies.


Roles of physical interactions in determining protein-folding mechanisms: molecular simulation of protein G and alpha spectrin SH3.
Protein-folding mechanisms of two small globular proteins, IgG binding domain of protein G and alpha spectrin SH3 domain are investigated via Brownian dynamics simulations with a model made of coarse-grained physical energy functions responsible for sequence-specific interactions and weak Gō-like energies. The folding pathways of alpha spectrin SH3 are known to be mainly controlled by the native topology, while protein G folding is anticipated to be more sensitive to the sequence-specific effects than native topology. We found in the folding of protein G that the C terminal beta hairpin is formed earlier and is rigid, once ordered, in the presence of an intact C terminal turn. The alpha helix is found to exhibit repeated partial formations/deformations during folding and to be stabilized via the tertiary contact with preformed beta sheets. This predicted scenario is fully consistent with experimental phi value data. Moreover, we found that the folding route is critically affected when the hydrophobic interaction is excluded from physical energy terms, suggesting that the hydrophobicity critically contributes to the folding propensity of protein G. For the folding of alpha spectrin SH3, we found that the distal beta hairpin and diverging turn are parts formed early, fully in harmony with previous results of simple Gō-like and experimental analysis, supporting that the folding route of SH3 domain is robust and coded by the native topology. The hybrid method provides useful tools for analyzing roles of physical interactions in determining folding mechanisms.


Wrist Deformity in Children and Adolescents with b-thalassemia on Oral Iron Chelation Therapy.
To describe a novel wrist deformity in b-thalassemia major patients, and their radiographic and magnetic resonance imaging findings. 30 patients with b-thalassemia major who were noticed to have ulnar deviation at wrist joint were evaluated for previous history of medications, serum ferritin levels, presence of pain and swelling at the wrist joint, and the duration of iron chelation therapy. Radiographs of wrist and limited magnetic resonance imaging (MRI) sequences were obtained in 30 and 15 patients, respectively. Radiographs revealed varying severity of distal ulnar shortening, distal radial slanting and presence of soft tissue distal to the ulna. MRI showed similar deformities along with abnormal marrow signal at distal ulnar ends; in 8 patients, a soft tissue distal to the distal end of ulna was noted. Varying severity of radiological abnormalities, predominantly affecting the distal ulna, are present in children and adolescents with b-thalassemia receiving oral chelation therapy.


A case of disseminated M. bovis bacillus Calmette-Guérin (BCG) disease after one month of BCG bladder infusion therapy and analysis of 77 cases of suspected BCG infection in Japan, 2017-2022.
Bacillus Calmette-Guerin (BCG) intravesical injections are used as adjuvant therapy for superficial bladder cancer. We report a case of a 78-year-old man who developed disseminated M. bovis BCG disease mimicking miliary tuberculosis early after BCG intravesical infusion. He started coughing after receiving three rounds of BCG for superficial bladder tumors, following transurethral resection of the tumors, approximately one month after initiation. Computerized tomography (CT) images showed diffuse nodular shadows in the bilateral lung fields with a random pattern. Consequently, disseminated BCG disease was diagnosed. Treatment with isoniazid, rifampicin, and ethambutol was initiated. Nine months after initiating treatment, CT showed the disappearance of the miliary shadows. We also discussed 77 cases of suspected BCG infection and the requests for Mycobacterium bovis BCG identification at our institution from 2017 to October 2022. Of these, 76 cases were M. bovis BCG, and 1 case was M. tuberculosis. Since M. tuberculosis can be identified in some patients with suspected BCG infection, it is crucial to distinguish between the two based on pathogenicity.


[Modification of a method to study cell fusion induced in vitro by viruses].
A studying method of viruses induced cell-fusion is described, using monolayers of Vero monkey cells. Nature and density of cells and infection multiplicity are studied. The sensibility is equal to 4 hemagglutining units of Sendaï and accuracy about +/- 3 % of polykaryocytosis. Quantification. comparison and kinetics study of cell-fusion are allowed by this method which may lead to systematic and routine exploration of cell-fusion ability of biological extracts from slow degenerative diseases of the human central nervous system.


On-chip dielectrophoretic recovery and detection of a lactate sensing probiotic from model human saliva.
Early detection has led to increased survival for multiple cancers; however, the 5-year survival rate of oral carcinoma (OC) has remained at 40% for the last several decades. Screening for OC is routinely done via visual examinations, followed by tissue biopsy and laboratory testing. Point-of-care testing would be a more convenient and widely available alternative for at-risk individuals. Increased lactate production is a hallmark of many head-and-neck tumors, due to the Warburg Effect, where tumor cells favor glycolysis in the place of oxidative phosphorylation. To detect excess lactate, we have modified the commensal bacterium Escherichia coli Nissle 1917 to express fluorescent reporter genes in response to extracellular lactate. Administering this commensal as a mouth wash and subsequently collecting saliva for the detection of the reporter may allow for noninvasive, early detection of cancerous lesions in at-risk individuals. Furthermore, we demonstrate a new on-chip electrokinetic technique to recover these probiotic probes from model saliva fluid to improve the detection of reporter gene activation.


Is the Choice of Cost-Effectiveness Threshold in Cost-Utility Analysis Endogenous to the Resulting Value of Technology? A Systematic Review.
Cost-utility analysis (CUA) is widely used for health technology assessment; however, concerns exist that cost-utility analysts may suggest higher cost-effectiveness thresholds (CETs) to compensate for technologies of relatively lower value. We explored whether selection of a CUA study's CET was endogenous to estimated incremental cost-effectiveness ratios (ICERs). We systematically reviewed the US cost-effectiveness literature between 2000 and 2017 where studies with explicit CET and ICERs were included. We classified the ratio of studies hypothesized to analyze cost-effective technologies at low CETs (i.e., less than $100,000/quality-adjusted life-year [QALY]) vs higher CETs (i.e., $100,000-$150,000/QALY) relative to their ICER, using a Chi square test to examine whether technologies that were cost effective at high CETs would still be cost effective at lower thresholds. We also performed fixed-effects linear regression exploring the associations between ICERs and reported CETs over time. Among 317 ICERs reviewed: (A) 185 had an ICER < $50,000/QALY; (B) 53 had $50,000 ≤ ICER, < $100,000; (C) 20 had $100,000 ≤ ICER < $150,000; and (D) 59 had an ICER ≥ $150,000. Chi square testing showed a strong association (p < 0.001) between estimated ICER values and chosen CET, illustrating a lack of independence between the two. The regression analysis indicated that CETs have a baseline value of $52,000 and grow by $0.37 for each dollar increase in the estimated ICER. Cost-effectiveness thresholds represent the hypothesis tests of typical CUAs. Our analysis highlights that most CUAs that cite high CETs also result in greater ICERs for the novel interventions that they investigate; thus, these interventions would otherwise not have been cost effective at lower CETs. Selection of a CET may come after the ICER is calculated to infer value that suits a hypothesis.


Acute blistering diseases on the burn ward: Beyond Stevens-Johnson Syndrome and Toxic Epidermal Necrolysis.
Stevens-Johnson Syndrome and Toxic Epidermal Necrolysis are on a spectrum of rare reactions primarily attributable to drugs. Timely diagnosis, cessation of the offending drug and burn center care are associated with favorable outcomes. Acute blistering disease has a wide differential diagnosis, including autoimmune bullous disease and other drug reactions. The aim of our study was to identify the final diagnosis in patients transferred for widespread blistering disease and to identify clinical features at admission predicting final diagnosis. We performed a 5-year retrospective chart review (2006-2011) of the clinical features at admission of patients transferred to a burn ward with widespread blistering disease. Clinical features at admission were compared between patients. 12 patients had a final diagnosis of Stevens-Johnson Syndrome or Toxic Epidermal Necrolysis and 7 patients had an alternative final diagnosis. Skin detachment surface area at admission was superior in the Stevens-Johnson Syndrome/Toxic Epidermal Necrolysis group. Presence of tense bullae and pustules was associated with an alternative final diagnosis. Extensive skin detachment surface and morphological features (tense bullae, pustules) were statistically significant clinical clues to final diagnosis. Patients transferred for widespread blistering disease should be thoroughly evaluated in order to exclude other causes of acute blistering disease.


Diclofenac sodium gel therapy as an alternative to actinic cheilitis.
Actinic cheilitis (AC) is a potentially malignant lesion caused by prolonged exposure to ultraviolet light. The aim of this research was to analyze the efficacy of diclofenac sodium 3% gel in the treatment of this condition, through clinical follow-up. Thirty-one patients diagnosed with AC were instructed to perform a topical application of the gel three times a day for a period of 90 days. In each visit, a digital photography was obtained for verified progress and response to treatment. Two researchers evaluated all images after treatment was completed and assigned the following scores regarding clinical aspect of the lip: 1, complete improvement; 2, partial improvement; 3, no changes; 4, worsening of the clinical condition. In addition, the patients' tolerability to the drug and their satisfaction after treatment were evaluated. Twelve cases abandoned the treatment for reasons unrelated to the study. Ten participants showed total remission of all clinical features of the lesion and three had partial improvement of the characteristics. One participant presented worsening of clinical condition, and in five cases, treatment was discontinued due to development of mild adverse effects at the site of gel application. Regarding satisfaction analyses and tolerability to the drug, from 14 patients who completed treatment without adverse effects or complications, most agreed fully that they were satisfied with the therapy (n = 11) and that the drug was not irritating to the mouth (n = 9). Patients are being monitored without clinical signs of recurrence and/or progression of the lesions. Topical application of the drug has provided a convenient and well tolerated in most cases. Diclofenac sodium gel (3%) may be a promising alternative for treatment of actinic cheilitis.


Social Determinants of Health and Mortality After Premature and Non-premature Acute Coronary Syndrome.
To describe and compare the determinants of 1-year mortality after premature vs non-premature acute coronary syndrome (ACS). Participants presenting with ACS were enrolled in a prospective registry of 29 hospitals in 4 countries, from January 22, 2012 to January 22, 2013, with 1-year of follow-up data. The primary outcome was all-cause 1-year mortality after premature ACS (men aged <55 years and women aged <65 years) and non-premature ACS (men aged ≥55 years and women aged ≥65 years). The associations between the baseline patient characteristics and 1-year mortality were analyzed in models adjusting for the Global Registry of Acute Coronary Events (GRACE) score and reported as adjusted odds ratio (aOR) (95% CI). Of the 3868 patients, 43.3% presented with premature ACS that was associated with lower 1-year mortality (5.7%) than those with non-premature ACS. In adjusted models, women experienced higher mortality than men after premature (aOR, 2.14 [1.37-3.41]) vs non-premature ACS (aOR, 1.28 [0.99-1.65]) (P interaction=.047). Patients lacking formal education vs any education had higher mortality after both premature (aOR, 2.92 [1.87-4.61]) and non-premature ACS (aOR, 1.78 [1.36-2.34]) (P interaction=.06). Lack of employment vs any employment was associated with approximately 3-fold higher mortality after premature and non-premature ACS (P interaction=.72). Using stepwise logistic regression to predict 1-year mortality, a model with GRACE risk score and 4 characteristics (education, employment, body mass index [kg/m2], and statin use within 24 hours after admission) had higher discrimination than the GRACE risk score alone (area under the curve, 0.800 vs 0.773; P comparison=.003). In this study, women, compared with men, had higher 1-year mortality after premature ACS. The social determinants of health (no formal education or employment) were strongly associated with higher 1-year mortality after premature and non-premature ACS, improved mortality prediction, and should be routinely considered in risk assessment after ACS.


Endocrine tumours of the gastrointestinal tract. Transplantation in the management of metastatic endocrine tumours.
Patients with neuroendocrine tumours often present with synchronous liver metastases or develop hepatic metastases in the course of their disease. A complete removal of liver metastases with an intention to cure may be accomplished by liver resection or, if hepatic disease is disseminated or hormonal symptoms and pain cannot be controlled medically, by total hepatectomy and transplantation. The indications for orthotopic liver transplantation for metastatic neuroendocrine tumour disease should be anchored in a multimodal and multidisciplinary therapeutic approach. Approximately, 120-130 cases of orthotopic liver transplantation for neuroendocrine tumours have been published so far, but follow-up after transplantation has been limited, and most reports comprise a small number of patients. After considering published studies and data, some recommendations may be given, although these are based on a low level of evidence. After excluding extrahepatic tumour manifestations by imaging procedures and diagnostic laparoscopy, the indication should be chosen restrictively. Few prognostic markers, for example age below 50 years and absence of concurrent extensive surgery, were identified by multivariate analysis in a large retrospective analysis. The prognostic impact of primary tumour localisation is still controversial. However, further indicators of favourable long-term prognosis are needed. Tumour biology characterised by Ki67 and E-cadherin expression may help to identify patients with a favourable outcome so that patient selection can be improved, but this needs further evaluation in larger patient cohorts. Orthotopic liver transplantation for patients with remission of disease or stable disease under medical treatment, and orthotopic liver transplantation for palliative reasons, should be restricted to selected individual cases.


Insulin-like growth factor-1 and insulin mediate transient site-selective increases in tau phosphorylation in primary cortical neurons.
The modulation of tau phosphorylation and localization in response to insulin-like growth factor-1 or insulin was examined in primary cultures of rat cortical neurons. Insulin and insulin-like growth factor-1 treatment resulted in a rapid and transient increase in tau phosphorylation at specific epitopes. These effects were completely inhibited by lithium, revealing that the insulin and insulin-like growth factor-1 induced changes in tau phosphorylation were mediated by glycogen synthase kinase-3beta. In addition, the increase in tau phosphorylation directly correlated with a transient dissociation of tau from the cytoskeleton, indicating that insulin and insulin-like growth factor-1 treatment resulted in a change in tau localization. Using immunocytochemistry, it was also demonstrated that treatment of neurons with insulin-like growth factor-1 for 3 min resulted in a redistribution of tau to the growth cone and the distal segment of the axons. Further, insulin-like growth factor-1 treatment resulted in an increased immunoreactivity with the phospho-dependent antibody AT8 in the same areas of the axons. Thus, the phosphorylation state and distribution of tau can be modulated by insulin and insulin-like growth factor-1 signaling pathways involving glycogen synthase kinase-3beta. We propose that by transiently increasing tau phosphorylation, insulin and insulin-like growth factor-1 may contribute to the reorganization of the cytoskeleton necessary for the development and growth of the neurites.


Role of the pharmacist on a general medicine acute care for the elderly unit.
The prevalence of prescription medication use increases with age, and older adults are at increased risk of harm from medication use. To describe the role of a pharmacist on a General Medicine Acute Care of the Elderly (GM-ACE) Unit. A job description was prepared, and a clinical pharmacist specializing in internal medicine was re-assigned to participate in multidisciplinary rounds on the ACE unit twice weekly and to work with a unit-based pharmacist assigned to multiple units. The clinical pharmacist also provided formal education on geriatric pharmacotherapy for other health care providers. Interventions were defined as changes in the medical record and provision of drug information. Interventions were tracked with an existing form and sorted by category. Data on interventions were presented to the pharmacy and therapeutics committee routinely. After 3 months, the clinical pharmacist accomplished 76 interventions in the areas of agent selection, dose optimization, addition of therapy, deletion of therapy, medication reconciliation, intravenous to oral conversion, technology improvements, and drug information/patient education. Expanding the role of the pharmacist in a GM-ACE unit has improved the medication use process in a high-risk population through improvements in medication overuse, medication underuse, dosing, medication reconciliation, patient education, and health care provider education.


Signs of sensory deprivation versus cerebral injury in post-hip-fracture patients.
Patients with a primary diagnosis of "postoperative hip fracture" often have a poor prognosis because of their inability to participate in rehabilitation programs. They tend to remain in a state of physical inactivity and sensory deprivation. The signs and symptoms of head injury associated with the hip fracture may be overlooked and attributed to sensory deprivation. Thirty-nine patients with fractured hip and 18 with multiple fractures were studied by means of electroencephalography, echoencephalography, and computer axial tomography. In 7, the findings indicated a cerebral lesion. At operation, a subdural hematoma was found in 5 patients and a tumor in 2; all but one made a dramatic recovery. Thus, after hip fracture, it is important that the patient be tested appropriately so that signs and symptoms of sensory deprivation may not be confused with those of an occult head injury. When indicated, surgical intervention may be a life-saving process.


Liver-infiltrating T lymphocytes cause hepatocyte damage by releasing humoral factors via LFA-1/ICAM-1 interaction in immunological liver injury.
To explore the mechanisms by which liver-infiltrating T lymphocytes cause hepatocyte damage in the liver injury induced by delayed-type hypersensitivity to picryl chloride. Nonparenchymal cells (NPC) were isolated 12 h after liver injury elicitation and fractionated into Kupffer cell-enriched (Fr. A) and lymphocyte-enriched populations (Fr. B). They were used as the effectors for coculture with hepatocytes. The cells in total NPC and Fr. B harvested at 12 h of liver injury were increased two- and six-fold respectively compared with those at 0 h. Fr. B, mainly including CD4+ and CD8+ T cells, exhibited a significantly stronger hepatotoxicity than total NPC did, while Fr.A did not. NPC at 12 h showed remarkably increased matrix metalloproteinase-2 and -9 activities indicative of infiltration potential through extracellular matrix. When NPC and hepatocytes were cultured in separated compartments in Transwell chamber, no hepatotoxicity was observed. However, 30 min-pre-contact with hepatocytes as stimulator significantly triggered NPC hepatotoxicity. The acquisition of such hepatotoxic potential was significantly abolished by anti-LFA-1 pretreatment for NPC or anti-ICAM-1 treatment for hepatocytes before contact. Both aprotonin and superoxide dismutase dose-dependently inhibited the hepatotoxicity. Liver-infiltrating T lymphocytes may be triggered by hepatocytes via LFA-1/ICAM-1 interaction to release toxic substances, such as proteases and oxygen radicals, which consequently lead to the hepatocyte damage.


The impact of primary repair timing on longitudinal visual outcomes after open globe injury.
The urgency with which to repair open globe injuries is a debated topic that lacks grounding in longitudinal visual outcomes data. We aim to test the association between primary repair timing and visual recovery potential following OGI. We performed a retrospective cohort study of medical records from a US academic medical center (7/2017 to 11/2021). We included all patients with a principal diagnosis of OGI, a documented date and time of injury, presentation, repair, and ≥ 3 months of complete follow-up data on visual outcomes. We excluded those with prior OGI in the same eye. We also tested the correlation of injury to repair time (ITR) in hours with best corrected visual acuity (BCVA, in logMAR units) at last follow-up in the general cohort and select subpopulations and the impact of repair delay on visual improvement over the follow-up period. One hundred twenty-nine patients with OGI were analyzed (91 patients with ≥ 3 months of follow-up). The majority were male (105/129, 81%) with a median age of 45 years. Most OGI involved zone 1 (57%), followed by zone 3 (24%), and zone 2 injuries (19%). Median ocular trauma score (OTS) was 60 (IQR 37-70); mean presenting BCVA was logMAR 1.9 (median 2.3, IQR 1.0-2.7). Median ITR was 22 h (IQR 15-30 h, range 5-199 h). ITR time did not significantly correlate with final BCVA (n = 91, β = - 0.003, 95% CI - 0.009-0.002, P = 0.233), nor did it significantly increase the odds of developing ocular complications or requiring secondary ocular surgeries (OR 0.985, 95% CI 0.967-1.002, P = 0.085). Additionally, the rate of BCVA improvement over subsequent months of follow-up did not significantly differ based on ITR time. Presenting BCVA (R2 = 0.701, P < 0.001) and OTS (R2 = 0.477, P < 0.001) significantly correlated with final BCVA, independent of repair delays. In this cohort of OGI patients, repair timing does not significantly correlate with final BCVA, and delays beyond 24 h do not significantly correlate with worse visual recovery potential. Repair time alone should be emphasized to a lesser extent as a prognosticator of visual potential, in favor of significant predictors such as the ocular trauma score and presenting visual acuity.


C3c concentrations in sera of persons living in areas with different levels of air pollution in Northrhine-Westphalia (Federal Republic of Germany).
A number of air pollutants is known to affect the immune system resulting in alterations of the complement activity in serum. We determined the levels of C3c, one of the activation products of the component C3, in serum samples obtained from various population groups living in areas with different levels of air pollution in Northrhine-Westphalia (FRG). The results show that the serum C3c levels tend to be increased in subjects living in areas with elevated levels of air pollution. By applying a linear regression model it can be demonstrated that a significant part of the variance of the C3c concentrations is explained by the residence area factor after making allowance for the effect of various confounders. The increase of the serum C3c concentrations appears to result from an increased activation of the complement component C3. The underlying mechanisms as well as the pathophysiological significance of these findings are unknown.


Metabolic effects of four intravenous nutritional regimens in patients undergoing elective surgery II.--Muscle amino acids and energy-rich phosphates.
Muscle amino acids and energy-rich phosphates have been determined in 28 patients before and after resection of a carcinoma of colon or rectum. Before operation the patients received a synthetic diet (Vivasorb for four days and postoperatively four different intravenous regimens with different amounts and proportions of amino acids were given for six days, the energy intake being the same for all groups. When compared with age-matched healthy controls the patients showed only slight increases in phenylalanine and isoleucine concentration in muscle and plasma with elevated plasma glutamate during the four days prior to the operation. Vivasorb treatment increased the concentration of alanine, glycine and methionine in both muscle and plasma while increases of threonine and histidine were seen only in muscle. Decreased concentrations of valine were found in muscle and plasma, while lysine and leucine decreased only in plasma. As an effect of operation, but independent of the nutritional regimen, muscle asparagine increased and muscle glutamine and glutamate decreased in all groups. The unique pattern of amino acid changes in postoperative trauma was confirmed in the present investigation but few differences in amino acid concentrations could be related to the various nutritional intakes. Alterations in the individual amino acid concentrations were not influenced by the intake of amino acids and seemed to have little relationship to the composition of any solution infused. Prior to operation there were low levels of ATP in muscle possibly due to immobilisation and chronic semistarvation but no changes in the energy rich phosphates could be attributed to trauma or nutrition. The content of muscle glycogen increased as a result of the Vivasorb supply indicating that the preoperative carbohydrate administration promotes muscle glycogen synthesis and postoperatively muscle glycogen was reduced significantly indicating enhanced glycogenolysis in the postoperative state. This finding emphasizes the importance of providing the patients with ample amount of glucose before and after surgery.


The influence of photoperiod on drought induction of dormancy in Lotus scoparius.
Many species of plants in Mediterranean climate regions have evolved deciduousness, causing reduced leaf area during the long summer drought characteristic of Mediterranean climates. This summer deciduous growth form has been considered a plant adaptation in Mediterranean regions allowing survival during periods of extreme water stress. Many studies have suggested the ecological importance of this growth form but few studies have examined the physiological stimulus for deciduousness.Previous data indicate that abscission in Lotus scoparius (a mediterranean California deciduous species) is influenced by both photoperiod and water stress in a complex manner. Here the physiological basis of long day enhanced leaf fall during water stress is investigated.Examination of water potential components indicate an osmotic adjustment with incresing water stress which enables the maintenance of turgor at lower water potentials. Osmotic adjustment in plants grown under long photoperiods was greater than that in short photoperiods. Therfore, long day enhanced abscission during water stress was not due to a greater susceptability to turgor loss during long days. Rather, long day treatment caused these plants to initiate dormancy (as indicated by soluble protein concentrations) during the onset of water stress. The dormant condition could not be released by subsequent release from water stress. Apparently, Lotus scoparius has evolved a photoperiodic control (presumably through growth regulators) over the initiation of dormancy during water stress. The adaptive significance of this photoperiodic control over the leaf abscission response to water stress relates to the variable climate of Mediterranean regions.


Randomized clinical trial of surgically assisted rapid maxillary expansion with two and three segments for nasal breathing.
To evaluate surgically assisted rapid maxillary expansion (SARME), with osteotomies separating the maxilla into two segments (SARME-2S) and three segments (SARME-3S), on obstruction symptoms and nasal cavity dimensions in patients with maxillary transverse skeletal deficiency (MTSD). Sixteen patients with MTSD of 7 mm or above were evaluated in each group, for a total of 32 patients. All patients were evaluated pre- and postoperatively up to 10 months after the expander activations. The minimum cross-sectional area (MCA) and the volume of the nasal cavities were identified by acoustic rhinometry. The Nasal Obstruction Symptom Evaluation (NOSE) scale questionnaire was applied. The palate surface area (PSA) was measured, via digitized maxillary models, as a criterion for comparison with the other variables studied. There was no difference between the groups (p = 0.370) and was verified a significant increase in PSA postoperatively. MCA showed a small increase without statistical significance, and together with the volume of the nasal cavities remained constant during the study. NOSE scale scores decreased significantly in the postoperative periods, implying a decrease in nasal obstruction symptoms in both groups. SARME with two and three segments show similar results, and both may improve nasal obstruction symptoms when present in patients with MTSD. SARME, regardless of the chosen surgical technique, should follow the recommendation to correct just the MTSD. While an improvement in nasal breathing is expected, this must be understood as likely, but not certain.


Occurrence of Crassulacean acid metabolism in Cissus trifoliata L. (Vitaceae).
Evidence for the operation of CAM in the deciduous climber, Cissus trifoliata L., was obtained in field and laboratory studies. Under natural conditions, diurnal oscillations of titratable acidity and colorimetric measurements of night CO2 fixation, determined for a period of two and a half years, suggested that acid accumulation was related to plant water status, assessed through the daily courses of stomatal resistance and xylem water potential during dry and rainy seasons. These findings were confirmed by gas exchange studies under controlled conditions which showed that the plant fixed all its CO2 during the day when it was well irrigated; as water stress increased, dark CO2 uptake gradually replaced fixation during the day until the plant only performed dark fixation. In severe water stress, even the rate of the latter process decreased until leaves fell.


Isolation of N-nitrosodimethylamine from drug substances using solid-phase extraction-liquid chromatography-tandem mass spectrometry.
N-Nitrosodimethylamine (NDMA) has been detected in some drug substances and pharmaceutical products containing sartans, ranitidine and metformin, and a potential risk of NDMA contamination exists in other drug substances and their pharmaceutical products. To quantitate NDMA in various drugs having diverse physicochemical properties, a specific, sensitive, and reliable analytical method is required, in addition to methods that can be applied to a class of nitrosamines. We aimed to develop an off-line isolation method for NDMA in drug substances using SPE for quantification with LC-APCI-MS/MS. Impediments to accurate quantitation of NDMA in drug substances using LC-MS/MS and insufficient durability of the system are attributed to the extremely large amounts of active pharmaceutical ingredients (APIs) in sample solutions in comparison to the trace amount of NDMA. A reduced retention of NDMA and/or decreased separation from other substances in LC, matrix effect in MS detection, and undesirable contamination of instruments with API and other substances may be occasionally encountered, all of which consequently result in deterioration of system performance and generation of unreliable data, even in the cases where a divert valve is configured between the column and ion source of the MS instrument. To address these problems, an off-line NDMA isolation methodology from APIs exhibiting diverse physicochemical properties, namely ranitidine hydrochloride (ranitidine), metformin hydrochloride (metformin), nizatidine, valsartan, and telmisartan, was developed. The applicability of the method was confirmed by batch analysis of metformin and ranitidine. Furthermore, contrary to previous reports, NDMA was found to be stable over a wide pH range. The proposed methodology and data from this study would contribute to the control of NDMA contamination in various drugs to realize the safe delivery of pharmaceuticals to patients.


Effects of bottom trawling on trace metal contamination of sediments along the submarine canyons of the Gulf of Palermo (southwestern Mediterranean).
Submarine canyons are preferential pathways for transport of particulate matter and contaminants from the shelf to the deep sea. The Gulf of Palermo continental margin has a very narrow shelf (about 2-3 km wide on average) and is incised by several submarine canyons that favour shelf-slope sediment transfer. A sediment core collected on the outer shelf and six sediment cores taken at different depths along the Oreto, Eleuterio and Anerella submarine canyons were analysed to study the transfer and historical record of trace metal contamination in the Gulf of Palermo continental margin. Trace metals, major elements, organic carbon and sediment grain size were analysed in these cores, which were dated with 210Pb to assess their historical compositional evolution since the late 19th century. Hg, Pb, Cu, Zn and Cd content increased until the 1970s and 1980s, associated with the increase in urbanization and industrial activities in the Palermo area, and Hg was the contaminant that reached the highest enrichments. However, the increasing trend of these metals contamination was reversed in the 1970s and 1980s, coinciding with drastic changes in the terrigenous content and grain size of sediments in the canyon axes. These changes occurred when bottom trawling fleets expanded to deeper fishing grounds equipped with powerful trawlers around the Gulf of Palermo canyon heads and flanks and along the Oreto canyon axis. Bottom trawlers have resuspended large amounts of sediment, which have been transferred into the canyons since the 1970s and 1980s and have thus increased sediment accumulation rates. This resuspended sediment has been mixing with the sediment transferred and accumulated along the canyons, diluting and reducing its trace metal contamination levels since the expansion of the bottom trawling fleets.


Immunoproteasome subunit PSMB8 regulates microglia-mediated neuroinflammation upon manganese exposure by PERK signaling.
Excessive manganese (Mn) exposure gives rise to various neurological disorders, including motor dysfunction and cognitive impairment. Microglia-mediated neuroinflammation plays an essential role in the pathogenesis of Mn neurotoxicity. However, the underlying mechanisms have not been fully clarified. Immunoproteasome is a specialized proteasome. Recent studies have shown that immunoproteasome, especially catalytic subunit PSMB8, is highly associated with various neurological diseases. Whether PSMB8 is involved in Mn-neurotoxicity is still unknown. In this study, in vivo and in vitro models were established, and our data showed that Mn exposure upregulated the expression and activity of PSMB8. Selective inhibition of PSMB8 mitigated neuroinflammation with reduced microglial activation and fewer TNF-α, iNOS, and CCL12 production in Mn-treated mice and BV2 cells. Learning and memory tests and Golgi staining further confirmed that inhibition of PSMB8 alleviated Mn-induced recognition memory impairments and synapse deficits. Besides, we found that blocking of PERK signaling inhibited Mn-induced elevation of PSMB8. And inhibition of PSMB8 reduced the phosphorylation of NF-κB p65. Together, our data demonstrated that PSMB8 played an essential role in microglia-mediated neuroinflammation upon Mn exposure, and the underlying mechanisms may be via PERK/NF-κB pathways. These results provide a novel target for the prevention and treatment of Mn-neurotoxicity.


Effect of the mono- and tetra-sialogangliosides, GM1 and GQ1b, on long-term potentiation in the CA1 hippocampal neurons of the guinea pig.
Effects of the mono- and tetra-sialogangliosides, GM1 and GQ1b, on long-term potentiation (LTP) were investigated in the CA1 neurons of guinea-pig hippocampal slices. The magnitude of LTP induced by a strong tetanus (100 Hz, 100 pulses) was not significantly affected by application of either ganglioside. In contrast, when LTP was induced by a weak tetanus (100 Hz, 4 pulses), a significantly greater LTP was induced in the presence of either ganglioside. Similarly, when slices were incubated in low-Ca2+ (1.0-1.1 mM) medium for more than 2 h, the LTP was usually small or absent, but showed a significant increase in amplitude of population spike (A-PS) when the slices were incubated with either GM1 or GQ1b (4-5 microg/ml). In addition, the application of GQ1b (4 microg/ml) reversed the blocking effect of an NMDA-receptor antagonist, APP-5 (10 microM), on the induction of LTP and resulted in forming LTP. Based on these findings, we conclude that GM1 and GQ1b exert positive modulatory effects on the induction of LTP in hippocampal CA1 neurons and suggest that GM1 and GQ1b may participate in the induction of LTP as donors of Ca2+ ions.


Perfluoroalkyl acids (PFAAs) in the aquatic food web of a temperate urban lake in East China: Bioaccumulation, biomagnification, and probabilistic human health risk.
The bioaccumulation and biomagnification of perfluoroalkyl acids (PFAAs) in temperate urban lacustrine ecosystems is poorly understood. We investigated the occurrence and trophic transfer of and probabilistic health risk from 15 PFAAs in the food web of Luoma Lake, a temperate urban lake in East China. The target PFAAs were widely distributed in the water (∑PFAA: 77.09 ± 9.07 ng/L), suspended particulate matter (SPM) (∑PFAA: 284.07 ± 118.05 ng/g dw), and sediment samples (∑PFAA: 67.77 ± 17.96 ng/g dw) and occurred in all biotic samples (∑PFAA: 443.27 ± 124.89 ng/g dw for aquatic plants; 294.99 ± 90.82 for aquatic animals). PFBA was predominant in water and SPM, with 40.11% and 21.35% of the total PFAAs, respectively, while PFOS was the most abundant in sediments (14.11% of the total PFAAs) and organisms (14.33% of the total PFAAs). Sediment exposure may be the major route of biological uptake of PFAAs. The PFAA accumulation capacity was the highest in submerged plants, followed by emergent plants > bivalves > crustaceans > fish > floating plants. Long-chain PFAAs were biomagnified, and short-chain PFAAs were biodiluted across the entire lacustrine food web. PFOS exhibited the greatest bioaccumulation and biomagnification potential among the target PFAAs. However, biomagnification of short-chain PFAAs was also observed within the low trophic-level part of the food web. Human health risk assessment indicated that perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA) posed health risks to all age groups, while the other PFAAs were unlikely to cause immediate harm to consumers in the region. This study fills a gap in the knowledge of the transfer of PFAAs in the food webs of temperate urban lakes.


Functional metagenomics reveals novel salt tolerance loci from the human gut microbiome.
Metagenomics is a powerful tool that allows for the culture-independent analysis of complex microbial communities. One of the most complex and dense microbial ecosystems known is that of the human distal colon, with cell densities reaching up to 10(12) per gram of faeces. With the majority of species as yet uncultured, there are an enormous number of novel genes awaiting discovery. In the current study, we conducted a functional screen of a metagenomic library of the human gut microbiota for potential salt-tolerant clones. Using transposon mutagenesis, three genes were identified from a single clone exhibiting high levels of identity to a species from the genus Collinsella (closest relative being Collinsella aerofaciens) (COLAER_01955, COLAER_01957 and COLAER_01981), a high G+C, Gram-positive member of the Actinobacteria commonly found in the human gut. The encoded proteins exhibit a strong similarity to GalE, MurB and MazG. Furthermore, pyrosequencing and bioinformatic analysis of two additional fosmid clones revealed the presence of an additional galE and mazG gene, with the highest level of genetic identity to Akkermansia muciniphila and Eggerthella sp. YY7918, respectively. Cloning and heterologous expression of the genes in the osmosensitive strain, Escherichia coli MKH13, resulted in increased salt tolerance of the transformed cells. It is hoped that the identification of atypical salt tolerance genes will help to further elucidate novel salt tolerance mechanisms, and will assist our increased understanding how resident bacteria cope with the osmolarity of the gastrointestinal tract.


Retinal detachment following phacoemulsification.
In a consecutive series of 1,500 eyes in which phacoemulsification had been performed, the incidence of subsequent retinal detachment was 3.6%. Vitreous loss occurred in 4.2% (63) of the 1,500 cases, and in this group of eyes the incidence of retinal detachment was 14% (9). The characteristics of retinal breaks and the configurations of retinal detachments following phacoemulsification were in general similar to cases following intracapsular surgery. The surgical reattachment rates in these eyes compared favorably to previous reports on the repair of retinal detachments.


Discovery of SARS-CoV-2 antiviral synergy between remdesivir and approved drugs in human lung cells.
SARS coronavirus 2 (SARS-CoV-2) has caused an ongoing global pandemic with significant mortality and morbidity. At this time, the only FDA-approved therapeutic for COVID-19 is remdesivir, a broad-spectrum antiviral nucleoside analog. Efficacy is only moderate, and improved treatment strategies are urgently needed. To accomplish this goal, we devised a strategy to identify compounds that act synergistically with remdesivir in preventing SARS-CoV-2 replication. We conducted combinatorial high-throughput screening in the presence of submaximal remdesivir concentrations, using a human lung epithelial cell line infected with a clinical isolate of SARS-CoV-2. This identified 20 approved drugs that act synergistically with remdesivir, many with favorable pharmacokinetic and safety profiles. Strongest effects were observed with established antivirals, Hepatitis C virus nonstructural protein 5A (HCV NS5A) inhibitors velpatasvir and elbasvir. Combination with their partner drugs sofosbuvir and grazoprevir further increased efficacy, increasing remdesivir's apparent potency > 25-fold. We report that HCV NS5A inhibitors act on the SARS-CoV-2 exonuclease proofreader, providing a possible explanation for the synergy observed with nucleoside analog remdesivir. FDA-approved Hepatitis C therapeutics Epclusa® (velpatasvir/sofosbuvir) and Zepatier® (elbasvir/grazoprevir) could be further optimized to achieve potency and pharmacokinetic properties that support clinical evaluation in combination with remdesivir.


Clonidine and guanfacine in hypertension.
Guanfacine, 1 to 3 mg/day, and clonidine, 0.1 to 0.3 mg twice a day, were compared in a 24-week double-blind, randomized, parallel study of 42 patients with hypertension that was inadequately treated by chlorthalidone, 25 mg/day. Mean reduction of blood pressure was 18/9 mm Hg after guanfacine and 14/8 mm Hg after clonidine. To determine the incidence of rebound hypertension, subjects were hospitalized for 7 days during chlorthalidone therapy for collection of baseline data and once again immediately after abrupt withdrawal of the alpha-agonist after 24 weeks of dosing. Although blood pressure and heart rate rose significantly in both groups, the changes after clonidine withdrawal were greater and occurred earlier (day 2) than those after guanfacine withdrawal (day 4). Forty percent of the subjects receiving guanfacine and 64% of subjects receiving clonidine had diastolic blood pressure elevations greater than or equal to 10 mm Hg from baseline. There were increases in urinary norepinephrine levels in both groups after drug withdrawal, but these correlated poorly with blood pressure rise. Side effects after guanfacine were much the same as those after clonidine. Guanfacine taken once a day provides an effective and safe alternative to clonidine in the management of essential hypertension.


Solubility of sulfur dioxide in tetraglyme-NH4SCN ionic liquid: high absorption efficiency.
An easily prepared ionic liquid was synthesized by a one-step method and applied in SO2 absorption efficiently. The cation of the ionic liquid is a supramolecular structure consisting of NH+ 4 and tetraglyme, similar to the structure of NH+ 4 and crown ether, and the anion is selected as SCN-. The ionic liquid has good thermal stability. Under the conditions of 293 K and 1 bar, one mol ionic liquid can absorb 2.73 mol SO2, which is about 30% higher than tetraglyme. The absorption mechanism was characterized using IR and NMR. And the results confirmed that the interaction mechanism between SO2 and the ionic liquid is a physical interaction rather than a chemical interaction.


A Handful of Sunscreen for Whole-Body Application.
The rule of thumb "Fill up a handful of sunscreen and spread it all over your body" has been used in several sun safety campaigns. The intention was to increase the applied sunscreen to obtain a quantity of 2 mg/cm2 to all accessible skin. The present study is the first to investigate how this advice works in practice, evaluated by quantity of sunscreen applied and amount of covered skin. Seventeen volunteers wearing swimwear were asked to "Fill up a handful and spread it all over your body." Before and after sunscreen application, the volunteers were photographed in black light. As sunscreen absorbs black light, the darkness of the skin increases with increasing amounts of applied sunscreen, making it possible to identify skin left without coverage. The sunscreen container was weighed before and after to quantify the amount of sunscreen applied. A median of 21% of the accessible skin was left completely without coverage. The 79% covered area was covered with a median of 1.12 mg/cm2, not the expected 2 mg/cm2. In practice, the advice "Fill up a handful of sunscreen and spread it all over your body" led to a better but still modest protection, compared to the intended effect.


Malignant entity of idiopathic ventricular fibrillation and polymorphic ventricular tachycardia initiated by premature extrasystoles originating from the right ventricular outflow tract.
The aim of this study was to assess the clinical characteristics and the efficacy of radiofrequency catheter ablation (RFCA) for idiopathic ventricular fibrillation (VF) and/or polymorphic ventricular tachycardia initiated by ventricular extrasystoles originating from the right ventricular outflow tract (RVOT). Ventricular fibrillation and/or polymorphic ventricular tachycardia are occasionally initiated by ventricular extrasystoles originating from the RVOT in patients without structural heart disease. Among 101 patients without structural heart disease in whom RFCA was conducted for idiopathic ventricular tachyarrhythmias arising from the RVOT, we examined the clinical characteristics and the efficacy of RFCA in 16 patients with spontaneous VF and/or polymorphic ventricular tachycardia initiated by the ventricular extrasystoles originating from the RVOT. Among 16 patients, spontaneous episodes of VF were documented in 5 patients, and 11 patients had prior episodes of syncope. Holter recordings showed frequent isolated ventricular extrasystoles with the same morphology as that of initiating ventricular extrasystoles, and non-sustained polymorphic ventricular tachycardia with short cycle length (mean of 245 +/- 28 ms) in all 16 patients. Radiofrequency catheter ablation by targeting the initiating ventricular extrasystoles eliminated episodes of syncope, VF, and cardiac arrest in all patients during follow-up periods of 54 +/- 39 months. Our data suggest that the malignant entity of idiopathic VF and/or polymorphic ventricular tachycardia was occasionally present in patients with idiopathic ventricular arrhythmias arising from the RVOT. Radiofrequency catheter ablation was effective as a treatment option for this entity.


Understanding allergic multimorbidity within the non-eosinophilic interactome.
The mechanisms explaining multimorbidity between asthma, dermatitis and rhinitis (allergic multimorbidity) are not well known. We investigated these mechanisms and their specificity in distinct cell types by means of an interactome-based analysis of expression data. Genes associated to the diseases were identified using data mining approaches, and their multimorbidity mechanisms in distinct cell types were characterized by means of an in silico analysis of the topology of the human interactome. We characterized specific pathomechanisms for multimorbidities between asthma, dermatitis and rhinitis for distinct emergent non-eosinophilic cell types. We observed differential roles for cytokine signaling, TLR-mediated signaling and metabolic pathways for multimorbidities across distinct cell types. Furthermore, we also identified individual genes potentially associated to multimorbidity mechanisms. Our results support the existence of differentiated multimorbidity mechanisms between asthma, dermatitis and rhinitis at cell type level, as well as mechanisms common to distinct cell types. These results will help understanding the biology underlying allergic multimorbidity, assisting in the design of new clinical studies.


Isolation of Desulfuromonas acetoxidans cytochrome c-551.5 from the mixed culture 'Chloropseudomonas ethylica'.
The mixed culture 'Chloropseudomonas ethylica' strain 2K has been grown on a medium which enhanced the yield of cytochrome c-551.5 from Desulfuromonas acetoxidans. The cytochrome was purified to homogeneity and an isoelectric point of 8.40 was determined. A determination of the amide content indicated that the cytochrome contains two more amides than previously reported.


Quantitative assessment of HR and NHEJ activities via CRISPR/Cas9-induced oligodeoxynucleotide-mediated DSB repair.
Homologous recombination (HR) and non-homologous end joining (NHEJ) are the two major mechanisms for the repair of DNA double-strand breaks (DSBs) in eukaryotic cells. Previously, we designed an assay for detecting NHEJ activity by using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, however, this approach cannot be used to predict the activity of HR repair. Hence, we developed a novel method that is capable of quantitatively measuring both HR and NHEJ activities via CRISPR/Cas9-induced oligodeoxynucleotide (ODN)-mediated DSB repair. In the present experimental procedures, the CRISPR/Cas9 plasmid was cotransfected with single-stranded ODN (ssODN) or blunt-ended double-stranded ODN (dsODN), both of which harbored a unique marker sequence. After the induction of site-specific DSBs by CRISPR/Cas9 system, the ssODN, functioned as the donor template for HR repair, could insert the marker sequence into the DSB sites, while the dsODN was embedded in the DSB sites through NHEJ pathway. Next, by means of PCR analysis using a specific primer for the marker sequence and the primers that flank the DSB sites, the relative amount of integrated marker sequence in the genomic DNA could be quantitatively determined. The correlation between the marker sequence abundance and the HR and NHEJ activities was confirmed by using the selective HR and NHEJ inhibitors. This accessible and rapid quantitative assay for HR and NHEJ activities might be useful for the future research of the DSB repair mechanisms.


Improving management of vaccine supply chains: A multi-methods evaluation of vSTEP in Zambia.
Equipping supply chain professionals at all levels of the health care systems with essential leadership and management skills is essential to achieving global immunization targets. Numerous and diverse professional development initiatives have been established to support vaccine supply chain managers in Gavi-eligible countries. However, COVID-19 catalyzed rapid innovation in approaches to support leadership development in times of rapid change, and systematic evaluation of these approaches can inform future workforce development strategies to ensure resilient health systems. Therefore, we sought to evaluate outcomes and short-term impact of the fully virtual Strategic Training Executive Program 2.0 (vSTEP 2.0) program in Zambia on participant engagement, skill development, and application of new skills to improve supply chain performance. We used a multi-method design to describe the outcomes and short-term impact of vSTEP 2.0 at three levels: (1) delegate response to the training experience (including participation, completion, and satisfaction); (2) delegate change in leadership skill (including achievement of learning objectives and change in competencies); and (3) application of new skills to impact supply chain performance. The program achieved high levels of engagement and an 83% graduation rate despite the length and rigor of the program. High satisfaction was evidenced by positive feedback from delegates on the quality and relevance of the program, especially in the context of COVID response (100% would "definitely" recommend). Delegates demonstrated significant growth in all domains of leadership competencies and applied their learnings to address a complex challenge in supply chain performance in their home organization. These findings demonstrate a strong appetite for leadership development support among vaccine supply chain professionals, especially in times of rapid change such as during the COVID response. These results also demonstrate the feasibility and impact of a fully virtual model for leadership development, with implications well beyond the COVID-19 pandemic.


Increase in mercury and methylmercury levels with depth in a fish assemblage.
Despite their remoteness, deep-sea species bioaccumulate mercury, mostly in the form of the neurotoxin methylmercury (MeHg). Although the concentration of MeHg in the water column is known to increase with depth down to a maximum found at the base of the permanent thermocline, the knowledge of the relationship between MeHg content in marine species and their depth of occurrence is limited. We analyzed total mercury (THg) and MeHg concentrations in 25 species of fish inhabiting the Avilés Submarine Canyon and its adjacent shelf (Cantabrian Sea, North-East Atlantic) between 50 and 1868 m depth. THg concentrations ranged from 0.03 μg g-1 in wet weight (ww) in Chauliodus sloani and 4.0 μg g-1 ww in Coryphaenoides guentheri. 65% of the species analyzed exceeded 0.5 μg g-1 ww of MeHg, the maximum level for safe consumption recommended by FAO/WHO. THg and MeHg contents in muscle tissue increased with the depth of occurrence of fish and was influenced by their habitat so that demersal species had higher THg content than pelagic species inhabiting the same depth. MeHg accounted for an average 76 ± 3.9% of THg (mean ± SD), which is lower than that reported for other fish communities and can be explained by the high concentration of Hg present in sediments of the Nalón estuary, which discharges right off the Avilés Canyon head. The % of THg as MeHg was also strongly correlated with δ15N values, confirming that MeHg can be an indicator of the trophic identity of a species within the food web.


Role of macrophage scavenger receptor 1 in the progression of dyslipidemia in acne vulgaris patients.
Macrophage scavenger receptor 1 gene (MSR1), is responsible for producing macrophage scavenger receptors. MSR1 is primarily located on the surfaces of various macrophage types and is known to exert a range of effects on the human body. These effects include influencing innate and adaptive immunological reactions, as well as contributing to the development of conditions such as atherosclerosis, dyslipidemia, liver and lung disease, and cancer. The unregulated assimilation of lipoproteins by MSR1 leads to the creation of macrophages rich in cholesterol that manifest as foam-like cells, ultimately contributing to dyslipidemia. This occurrence highlights the significance of MSR1 as a key player in the pathophysiology of dyslipidemia. In this study, we aimed to estimate variation in lipid profile in acne vulgaris (AV) patients. Also, we aimed to investigate the role of MSR1 in lipid profile variation. A case-control study consisting of 100 patients with AV and 104 healthy controls. Lipid profiles were assessed using normalized enzymatic processes and genotype analyses were performed by a polymerase chain reaction and standard Sanger sequencing. Predictions of variant effects were performed using in silico tools. Our results indicated that the levels of lipid profile were higher in patients with AV than in healthy patients. The two haplotypes that were most prevalent in the patients were TCAC (16.5%) and CAGG (15.47%), whereas the two haplotypes that were more prevalent in the controls were TAAC (16.43%) and CCAC (15.62%). IVS5.59 C > A and rs433235 A > G are in linkage disequilibrium. Additionally, rs433235 A > G has a significant linkage disequilibrium with rs3747531 C > G. In silico analysis, tools indicated that the rs433235 A > G variant was disease-causing. Patients diagnosed with TCAC and CAGG exhibited a higher prevalence compared to healthy patients with TAAC and CCAC. The linkage disequilibrium between rs433235 A > G and IVS5.59 C > A has been established. Furthermore, there appears to be significant linkage disequilibrium between rs3747531 C > G and rs433235 A > G. These findings support the notion that genetic variations may play a critical role in the pathogenesis of these conditions.


IDH-wild type glioblastomas featuring at least 30% giant cells are characterized by frequent RB1 and NF1 alterations and hypermutation.
Giant cell glioblastoma (GC-GBM) is a rare variant of IDH-wt GBM histologically characterized by the presence of numerous multinucleated giant cells and molecularly considered a hybrid between IDH-wt and IDH-mutant GBM. The lack of an objective definition, specifying the percentage of giant cells required for this diagnosis, may account for the absence of a definite molecular profile of this variant. This study aimed to clarify the molecular landscape of GC-GBM, exploring the mutations and copy number variations of 458 cancer-related genes, tumor mutational burden (TMB), and microsatellite instability (MSI) in 39 GBMs dichotomized into having 30-49% (15 cases) or ≥ 50% (24 cases) GCs. The type and prevalence of the genetic alterations in this series was not associated with the GCs content (< 50% or ≥ 50%). Most cases (82% and 51.2%) had impairment in TP53/MDM2 and PTEN/PI3K pathways, but a high proportion also featured TERT promoter mutations (61.5%) and RB1 (25.6%) or NF1 (25.6%) alterations. EGFR amplification was detected in 18% cases in association with a shorter overall survival (P = 0.004). Sixteen (41%) cases had a TMB > 10 mut/Mb, including two (5%) that harbored MSI and one with a POLE mutation. The frequency of RB1 and NF1 alterations and TMB counts were significantly higher compared to 567 IDH wild type (P < 0.0001; P = 0.0003; P < 0.0001) and 26 IDH-mutant (P < 0.0001; P = 0.0227; P < 0.0001) GBMs in the TCGA PanCancer Atlas cohort. These findings demonstrate that the molecular landscape of GBMs with at least 30% giant cells is dominated by the impairment of TP53/MDM2 and PTEN/PI3K pathways, and additionally characterized by frequent RB1 alterations and hypermutation and by EGFR amplification in more aggressive cases. The high frequency of hypermutated cases suggests that GC-GBMs might be candidates for immune check-point inhibitors clinical trials.


Stimulation with N-methyl-D-aspartate or kainic acid increases cyclic guanosine monophosphate-like immunoreactivity in turtle retina: involvement of nitric oxide synthase.
In brain and retina, stimulation with excitatory amino acids (EAA) can generate nitric oxide (NO) and increase levels of cyclic guanosine monophosphate (cGMP). Because nitric oxide synthase (NOS) has been found in retinas of all species examined to date, an NO signal-transduction pathway is likely to be present in all retinas. We tested the hypothesis that stimulation of ionotropic glutamate receptors in turtle retina would result in increases in cGMP through an NOS/NO/cGMP pathway. Following in vitro incubations of turtle eye cups with the glutamate receptor agonists, N-methyl-D-aspartate (NMDA) or kainic acid (KA), we quantified the increases in cGMP-like immunoreactivity (cGMP-LI) by using enzyme-linked immunosorbant assay (ELISA) and localized the increased cGMP-LI by using an antibody against cGMP. Stimulation with NMDA or KA increased cGMP-LI in bipolar and amacrine cells as well as in some somata in the ganglion cell layer. Either KA or NMDA produced statistically significant increases in total retinal cGMP-LI by ELISA. To test the involvement of NO, we used the NOS inhibitors 7-nitroindazole and L-nitroarginine. Both inhibitors blocked virtually all of the KA- or NMDA-stimulated increases in cGMP-LI. These results indicate that activation of ionotropic glutamate receptors can increase cGMP in select retinal neurons. Differences between the agonist-evoked increases of retinal cGMP-LI suggest that there can be specificity in the activation of the NOS/NO/cGMP signal-transduction pathway by glutamate. This suggests that, in addition to short-term electrical changes, activation of ionotropic glutamate receptors also may produce longer term modulatory or metabolic effects involving NO/cGMP.


Sensitive colorimetric detection of Listeria monocytogenes based on isothermal gene amplification and unmodified gold nanoparticles.
Listeria monocytogenes (L. monocytogenes), one of most problematic food-borne bacteria, is mainly transmitted through the food chain and may cause listeriosis. Therefore, the development of rapid and sensitive L. monocytogenes detection technique has become an urgent task. In this study, we proposed a method using hyperbranching rolling circle amplification (HRCA) combined with gold nanoparticle (GNP) based colorimetric strategy to offer an isothermal, highly sensitive and specific assay for the detection of L. monocytogenes. First, a linear padlock probe targeting a specific sequence in the hly gene was designed and followed with a ligation by Taq DNA ligase. After ligation, further amplification by HRCA with a thiolated primer and an unlabeled primer is performed. The resulting thiolated HRCA products were then captured onto GNP surface and made GNP more salt-tolerant. Detection of the bacteria can be achieved by a facilitated GNP based colorimetric testing using naked eyes. Through this approach, as low as 100 aM synthetic hly gene targets and about 75 copies of L. monocytogenes can be detected. The specificity is evaluated by distinguishing target L. monocytogenes from other bacteria. The artificial contaminated food samples were also detected for its potential applications in real food detection. This method described here is ideal for bacteria detection due to its simplicity and high sensitivity.


Effect of radiofrequency on patellar ligament repair of Wistar rats.
This study aimed to evaluate the effects of radiofrequency (RF) on patellar ligament repair through the analysis of type I and III collagens and immunostaining for TGF-β3. To evaluate the effect of RF on patellar ligament repair of Wistar rats, cross-sectional incision (60% of the width - grade I) was performed in patellar ligaments of the groups: lesion (L, n = 7), treated with RF on the 5-day (5RF, n = 7) and 7-day (7RF, n = 7) post injury were compared to control group (C, n = 7). Histological evaluation, immunohistochemistry, morphometry and statistical analysis were performed. At 10 days post injury, ligament rupture were observed only in L. Active fibroblasts, type 3 collagen and TGF-β3 in L, 5RF and 7RF was significantly (p < 0.05) higher than control (C). Type 1 collagen was significantly (p < 0.05) higher in C than L, 5RF and 7RF. A positive correlation (p < 0.05) was observed: TGF-β3 vs active fibroblasts and TGF-β3 vs type 3 collagen; otherwise, negative correlation (p < 0.05): type I collagen vs TGF-β3. These results suggest that RF seemed to accelerate the wound healing process of the patellar ligament and may be used as a non-invasive treatment of partial ligament injuries.


Characterization of outcomes of amino acid modifications using a combinatorial approach to reveal physical and structural perturbations: A case study using trastuzumab biosimilar.
Biopharmaceuticals, such as monoclonal antibodies, are considered as life-saving drugs for autoimmune diseases, cancer and infectious diseases. However, biotherapeutics tend to undergo chemical degradation during various stages of manufacturing. The conditions of chemical degradation, along with the physical degradation pathways, have a direct influence on the overall stability, safety and efficacy of these therapeutics. While site-specific chemical changes have been well-explored and investigated using various analytical approaches, the resulting conformational and structural changes have not been much studied. Thus, we explored various biophysical techniques for assessing the influence of three representatives forced degradation conditions viz. oxidation, deamidation, and glycation, in a model therapeutic trastuzumab biosimilar. The site-specific modifications caused by these stress conditions were analysed using high resolution mass spectrometry. While their thermodynamic and conformational consequences were investigated by using differential scanning colorimetry (Nano-DSC), circular dichroism (CD) spectroscopy, analytical ultracentrifugation (AUC), and dynamic light scattering (DLS). The investigated stress conditions resulted in reduced thermodynamic stability of mAb, as confirmed using Nano-DSC. Secondary structure analysis performed with CD spectroscopy indicated detectable structural alterations in the beta sheets of stressed samples. DLS and SV-AUC studies demonstrated an enhanced level of aggregation and fragmentation in presence of all stress conditions. Thus, the biophysical analytical toolkits, when used simultaneously, could offer deeper insights into the subtle conformational changes that result from site-specific chemical modifications in mAbs. Hence, these analytical approaches may serve as significant additions to the battery of techniques used for forced degradation analysis of biopharmaceuticals.


Association between perceived stress and hypertension in Chinese adults: a population-based survey.
It still remains unclear whether perceived stress is associated with hypertension. We aimed to investigate the association between perceived stress with hypertension in Chinese adults. Data on 10,823 participants aged 18 years or older were obtained from the 2015 China Health and Nutrition Survey (CHNS). Hypertension was defined as a mean systolic blood pressure ≥140 mmHg, diastolic blood pressure ≥90 mmHg, self-reported physician diagnosis of hypertension, and/or use of antihypertensive medications. Perceived stress was assessed by the 14-item perceived stress scale, with the scale data then divided into quartiles. Logistic regression models and restricted cubic spline regression were performed to evaluate the association between perceived stress and hypertension. After adjustment for age, gender, marital status, education level, per capita household income, employment status, residential areas, current smoking and alcohol consumption, the odds ratio (OR) [95% confidence interval (CI)] for every standard deviation (SD) increment in perceived stress was 0.94 (0.90-0.98); compared to the lowest quartile of perceived stress (Q1), the ORs (95% CIs) for hypertension were 0.88 (0.78-1.01) for Q2, 0.91 (0.81-1.03) for Q3, and 0.82 (0.72-0.92) for Q4 (P for trend < 0.001). The inverse association between perceived stress and hypertension was persistent in the subgroup analyses stratified by age, gender, employment status, residential areas, smoking status, and alcohol consumption. In addition, the restricted cubic spline curve revealed a linear and negative association between perceived stress and hypertension (P for nonlinearity = 0.872). Because this study was a cross-sectional design, future prospective studies are needed to confirm these findings.


Biotransport phenomena in freezing mammalian oocytes.
Water transport across the cell plasma membrane and intracellular ice formation (IIF)-the two biophysical events that may cause cell injury during cryopreservation-were studied by cryomicroscopy and modeling using mammalian (Peromyscus) oocytes. Unusually high activation energy for water transport across the cell plasma membrane was identified indicating that the water transport process is unusually sensitive to temperature (and cooling rate). Although literally all studies on IIF were conducted using protocols with ice-seeding (seeding extracellular ice usually at ≥-7 °C), it is not used for cell cryopreservation by vitrification that is becoming increasingly popular today. In this article, we show that ice-seeding has a significant impact on IIF. With ice-seeding and cooling at 60 °C/min, IIF was observed to occur over a wide range from approximately -8 to -48 °C with a clear change of the ice nucleation mechanism (from surface- to volume-catalyzed nucleation) at approximately -43 °C. On the contrary, without ice-seeding, IIF occurred over a much narrower range from approximately -19 to -27 °C without a noticeable change of the nucleation mechanism. Moreover, the kinetics of IIF without ice-seeding was found to be strongly temperature (and cooling rate) dependent. These findings indicate the importance of quantifying the IIF kinetics in the absence of ice-seeding during cooling for development of optimal vitrification protocols of cell cryopreservation.


A systematic review of cognitive performance in patients with childhood craniopharyngioma.
Craniopharyngiomas are rare brain tumors of the sellar/suprasellar region, often adversely affecting patients' physical and psychosocial functioning. Until a few years ago, knowledge on cognitive deficits in craniopharyngioma patients was based on little valid evidence, with considerable inconsistencies across studies. Findings from recent research, with partly larger sample sizes, add to existing evidence to provide a more clear and reliable picture. The current review aims to summarize and systemize current findings on cognitive deficits in childhood craniopharyngioma, taking account of patient- and treatment-related variables where possible. Those studies were included that reported results of childhood craniopharyngioma patients tested with formalized neuropsychological tests (irrespective of their age at study, group size ≥10). A systematic assignment of test results to subcomponents of broader cognitive domains (e.g. to specific memory systems and processes) allows for a first comprehensive overview of patterns of spared and impaired cognitive functions. We show that episodic memory recall in particular is impaired, largely sparing other memory components. In accordance with recent knowledge on mammillary function, patients with hypothalamic involvement appear to be at particular risk. Deficits in higher cognitive processes, relying on the integrity of the prefrontal cortex and its subcortical pathways, may also occur, but results are still inconsistent. To gain deeper insight into the pattern of deficits and their association with patient- and treatment-related variables, further multi-site research with larger cohorts is needed.


A field and laboratory study on factors affecting polymorphism in the rotifer Keratella tropica.
We present data on polymorphism of the rotifer Keratella tropica from an outdoor, mesoscale experiment and a series of laboratory induction assays. In the outdoor experiment the biomass of planktonic crustaceans was noticeably depressed by larval fish predation, and associated with this depression K. tropica underwent a striking reduction of caudal spines. Subsequent laboratory studies showed that the positive association between crustacean biomass and caudal spine length was a cause and effect relationship. This is the first record of rotifer morphological change as an indirect effect of fish predation. In laboratory experiments filtrates of monospecific cultures of a cyclopoid copepod, a calanoid copepod and 5 cladocerans induced a remarkable spine development. Morphological induction showed a direct relationship with the concentration of crustaceans, both under field and laboratory conditions. Long spines were found to be strong deterrents against small predators (Acanthocyclops robustus copepodites), but were useless against large ones (females of the same species). The morphotypes of K. tropica obtained by experimental induction from a single clone encompass much of the worldwide variation of the species.


Potent double prodrug forms of synthetic phosphoantigens.
Phosphoantigens are ligands of BTN3A1 that stimulate anti-cancer functions of γδ T cells, yet the potency of natural phosphoantigens is limited by low cell permeability and low metabolic stability. Derivatives of BTN3A1 ligand prodrugs were synthesized that contain an acetate-protected allylic alcohol and act as doubly protected prodrugs. A novel set of phosphonates, phosphoramidates, and phosphonamidates has been prepared through a new route that simplifies synthesis and postpones the point of divergence into different prodrug forms. One of the new prodrugs, compound 11, potently stimulates γδ T cell proliferation (72 h EC50 = 0.12 nM) and interferon γ response to loaded leukemia cells (4 h EC50 = 19 nM). This phosphonamidate form was > 900x more potent than the corresponding phosphoramidate, and the phosphonamidate form was also significantly more stable in plasma following acetate hydrolysis. Therefore, prodrug modification of phosphonate butyrophilin ligands at the allylic alcohol can both facilitate chemical synthesis and improve potency of γδ T cell stimulation.


Biochemical and hormonal parameters of lambs using guava (Psidium guajava L.) agro-industrial waste in the diet.
The objective of this research was to evaluate the biochemical and hormonal parameters of Santa Inês sheep that received diets with different levels of guava agro-industrial waste (GAW). Forty non-castrated male animals of the Santa Inês breed were used, with an average initial weight of 21.3 ± 2.6 kg, and an average age of 120 days. The animals were distributed in a completely randomized design, with five treatments and eight replicates, and fed with increasing GAW levels replacing hay (0.0, 7.5, 15.0, 22.5 and 30.0%). After the beginning of the diet period, blood samples were collected every 2 weeks approximately 3 h after feeding. Cholesterol concentration had a significant effect (P < 0.01) with the inclusion of GAW. The animals that ingested the control diet had the highest concentrations of cholesterol. T3 and T4 hormones showed a significant difference (P < 0.01) among the treatments, i.e. the animals that ingested the control diet had the lowest concentrations of these hormones. We concluded that the inclusion of GAW in up to 30% decreases the concentration of cholesterol and increases the concentrations of T3 and T4 in the blood, aiding the metabolism of the animals.


Pregabalin as an effective treatment for acute postoperative pain following spinal surgery without major side effects: protocol for a prospective, randomized controlled, double-blinded trial.
Patients experience considerable postoperative pain after spinal surgery. As the spine is located at the centre of the body and supports body weight, severe postoperative pain hinders upper body elevation and gait which can lead to various complications, including pulmonary deterioration and pressure sores. It is important to effectively control postoperative pain to prevent such complications. Gabapentinoids are widely used as preemptive multimodal analgesia, but their effects and side effects are dose-dependent. This study was designed to examine the efficacy and side effects of varying doses of postoperative pregabalin for the treatment of postoperative pain after spinal surgery. This is a prospective, randomized controlled, double-blind study. A total of 132 participants will be randomly assigned to the placebo (n = 33) group or to the pregabalin 25 mg (n = 33), 50 mg (n = 33), or 75 mg (n = 33) groups. Each participant will be administered placebo or pregabalin once prior to surgery and every 12 h after surgery for 72 h. The primary outcome will be the visual analogue scale pain score, total dose of administered intravenous patient-controlled analgesia, and frequency of rescue analgesic administered for 72 h from arrival to the general ward after surgery, subdivided into four periods: 1-6 h, 6-24 h, 24-48 h, and 48-72 h. The secondary outcomes will be the incidence and frequency of nausea and vomiting due to intravenous patient-controlled analgesia. Safety will be assessed by monitoring the occurrence of side effects such as sedation, dizziness, headache, visual disturbance, and swelling. Pregabalin is already widely used as preemptive analgesia and, unlike nonsteroidal anti-inflammatory drugs, is not associated with a risk of nonunion after spinal surgery. A recent meta-analysis demonstrated the analgesic efficacy and opioid-sparing effect of gabapentinoids with significantly decreased risks of nausea, vomiting, and pruritus. This study will provide evidence for the optimal dosage of pregabalin for the treatment of postoperative pain after spinal surgery. ClinicalTrials.gov NCT05478382. Registered on 26 July 2022.


Energy expenditure of southern right whales varies with body size, reproductive state and activity level.
Quantifying the energy expenditure of animals is critical to understanding the cost of anthropogenic disturbance relative to their overall energy requirements. We used novel drone focal follows (776 follows, 185 individuals) and aerial photogrammetry (5372 measurements, 791 individuals) to measure the respiration rate and body condition loss of southern right whales (Eubalaena australis) on a breeding ground in Australia. Respiration rates were converted to oxygen consumption rate and field metabolic rate (FMR) using published bioenergetic models. The intra-seasonal loss in body condition of different reproductive classes (calves, juveniles, adults, pregnant and lactating females) was converted to blubber energy loss and total energy expenditure (TEE). Using these two metrics, we tested the effects of body size, reproductive state and activity level on right whale energy expenditure. Respiration rates and mass-specific FMR decreased exponentially with an increase in body size, as expected based on allometric scaling. FMR increased curvilinearly with an increase in swim speed, probably as a result of increased drag and increased locomotion costs. Respiration rates and FMR were 44% higher for pregnant and lactating females compared with those of adults, suggesting significant costs of fetal maintenance and milk production, respectively. The estimated FMR of adults based on their respiration rates corresponded well with the estimated TEE based on body condition loss. The rate of decline in body condition of pregnant and lactating females was considerably higher than expected based on respiration rates, which probably reflects the milk energy transfer from mothers to calves, which is not reflected in their FMR.