Unravelling the influence of surface lipids on the structure, dynamics and interactome of high-density lipoproteins.
Surface lipids influence the biological activities of high-density lipoproteins (HDLs) but their species-specific effects on HDL structure, dynamics, and surface interactome has remained unclear. Building upon the five-lipid species HDL models developed and characterised in previous work, representative models of the major HDL subpopulations found in human plasma containing apolipoprotein A-I (apoA-I) have been studied using molecular dynamics simulation to describe their varying degrees of surface lipidome complexity. Specifically, two additional sets of representative HDL subpopulation particles were developed, one with sphingomyelin (SM) and the other with SM, phosphatidylethanolamine, phosphatidylinositol, and ceramide in quantities reflecting average levels characterised for HDL subpopulations derived from normolipidemic patients. These lipid species were assessed in terms of HDL size, morphology, dynamics, and overall interactome. The findings reveal that the presence of a representative SM fraction marginally enhanced HDL interfacial curvature and surface monolayer rigidity, manifesting in tighter phospholipid packing and slower surface lipid dynamics relative to SM-deficient HDL models. Furthermore, the presence of SM resulted in a reduction in the solvent exposure of core lipids and cholesterol molecules, whilst also enhancing apolipoprotein conformational flexibility and its overall twisting across the HDL surface. The hydrophobicity of apoA-I-bound lipid patches and the proportion of apoA-I hydrophobic surface area is enhanced by the overall lipidation of apoA-I irrespective of lipid composition. These findings offer new insights into how the surface lipid composition of different HDL subpopulations can significantly impact the overall interactome of HDL particles, potentially influencing subpopulation-specific biological functions like lipid scavenging and receptor interactions.


Association between treatment failure and hospitalization after receipt of neutralizing monoclonal antibody treatment for COVID-19 outpatients.
Neutralizing monoclonal antibodies (mAbs) are highly effective in reducing hospitalization and mortality among early symptomatic COVID-19 patients in clinical trials and real-world data. While resistance to some mAbs has since emerged among new variants, characteristics associated with treatment failure of mAbs remain unknown. This multicenter, observational cohort study included patients with COVID-19 who received mAb treatment between November 20, 2020, and December 9, 2021. We utilized electronic health records from a statewide health system plus state-level vaccine and mortality data. The primary outcome was mAb treatment failure, defined as hospitalization or death within 28 days of a positive SARS-CoV-2 test. COVID-19 mAb was administered to 7406 patients. Hospitalization within 28 days of positive SARS-CoV-2 test occurred in 258 (3.5%) of all patients who received mAb treatment. Ten patients (0.1%) died within 28 days, and all but one were hospitalized prior to death. Characteristics associated with treatment failure included having two or more comorbidities excluding obesity and immunocompromised status (adjusted odds ratio [OR] 3.71, 95% confidence interval [CI] 2.52-5.56), lack of SARS-CoV-2 vaccination (OR 2.73, 95% CI 2.01-3.77), non-Hispanic black race/ethnicity (OR 2.21, 95% CI 1.20-3.82), obesity (OR 1.79, 95% CI 1.36-2.34), one comorbidity (OR 1.68, 95% CI 1.11-2.57), age ≥ 65 years (OR 1.62, 95% CI 1.13-2.35), and male sex (OR 1.56, 95% CI 1.21-2.02). Immunocompromised status (none, mild, or moderate/severe), pandemic phase, and type of mAb received were not associated with treatment failure (all p > 0.05). Comorbidities, lack of prior SARS-CoV-2 vaccination, non-Hispanic black race/ethnicity, obesity, age ≥ 65 years, and male sex are associated with treatment failure of mAbs.


The Addition of Topotecan to Melphalan in the Treatment of Retinoblastoma with Intra-arterial Chemotherapy.
To evaluate the effect of adding topotecan to melphalan for the treatment of retinoblastoma using intra-arterial chemotherapy (IAC). Single-center, consecutive case series. All eyes treated with IAC at the University of Siena, Siena, Italy, from 2008 to 2019. Eyes were treated via IAC with either melphalan monotherapy or melphalan plus topotecan. The characteristics and outcomes of these 2 groups were compared. The main outcome measure was globe salvage rate. Additionally, a complete summary of all adverse events for all eyes was compared between groups and included local, regional, and systemic events causing both transient and permanent effects. A total of 193 patients and 208 eyes were treated with IAC between April 2008 and October 2019. Melphalan alone (MA) was used to treat 44 patients and 50 eyes for a total of 191 procedures. The combination of melphalan plus topotecan (MPT) was used to treat 149 patients and 158 eyes for a total of 780 procedures. Groups were similar in terms of age at presentation. The MPT group included more advanced eyes (P < 0.001) and had shorter follow-up time (mean 47 vs. 120 months in the MA group, P < 0.001). The MPT group required less laser and cryotherapy after treatment (32% of eyes vs. 50% of eyes in the MA group, P < 0.001); there was no other difference in the number of adjuvant treatments required between groups. There was no difference in the number of acute adverse events, both systemic and local, between groups. There was no difference in the number of transient or permanent intraocular side effects between groups. Kaplan-Meier survival analysis estimated a better globe salvage rate in the MPT group (66%) compared with the MA group (58%, P = 0.05). In this case series, the addition of topotecan to melphalan did not alter the IAC side effect profile and may contribute to improved globe salvage.


Differential diagnosis of pancreatobiliary carcinoma from autoimmune pancreatitis-related diseases: a report of three cases.
"Japanese clinical guidelines for autoimmune pancreatitis" advised to carefully differentiate between two conditions: autoimmune pancreatitis (AIP) and associated sclerosing cholangitis (SC), and pancreatobiliary malignancy. We report a series of three cases for which differential diagnosis of pancreatobiliary carcinoma from AIP and associated SC was crucial. Three patients presented with biliary stenosis secondary to pancreatic swelling or mass lesion, followed by further examinations: Case 1 was first diagnosed as having tumor-forming pancreatitis associated with AIP but eventually proven to be pancreatic head carcinoma; case 2 was operated for suspected bile duct cancer combined with AIP and associated cholangitis, and early cancer was found in the resected specimen; case 3 was operated on for presumed cholangiocarcinoma combined with AIP-associated SC, but no malignancy was found. Current series of cases would raise an alert on diagnosis of AIP and associated SC, and pancreatobiliary malignancy should be carefully excluded by any means. Surgical intervention would be required in selected cases of this clinical entity.


Rhythmicity matters: Circadian and ultradian patterns of HPA axis activity.
Oscillations are a fundamental feature of neural and endocrine systems. The hypothalamic-pituitary-adrenal (HPA) axis dynamically controls corticosteroid secretion in basal conditions and in response to stress. Across the 24-h day, HPA axis activity oscillates with both an ultradian and circadian rhythm. These rhythms have been shown to be important for regulating metabolism, inflammation, mood, cognition and stress responsiveness. Here we will discuss the neural and endocrine mechanisms driving these rhythms, the physiological importance of these rhythms and health consequences when they are disrupted.


Impact of Ruptured Aneurysm Circulation on Mortality: A Nationwide Inpatient Sample Analysis.
This study investigates the effect of aneurysm circulation on mortality and patient outcomes after aneurysmal subarachnoid hemorrhage (SAH) within the United States. A retrospective cohort study was conducted using the Nationwide Inpatient Sample (NIS), a part of the Healthcare Cost and Utilization Project (HCUP), with ICD-10 codes for non-traumatic SAH between 2015-2016. Aneurysms were stratified as either anterior or posterior circulation. Multivariate logistic regression was used to find the impact of selected variables on the odds of mortality. The NIS reported 1,892 cases of non-traumatic SAH within the study period that were predominantly anterior circulation (82.6%), female (68.6%), white (57.7%), with mean age of 59.07 years, and in-hospital mortality of 21.4%. Anterior circulation aneurysms were associated with lower severity of initial illness (p = 0.014) but higher likelihood of vasospasm (p = 0.0006) than those of the posterior circulation. In a multivariate logistic regression analysis, mortality was associated with posterior circulation aneurysms (OR: 1.42; CI 95% 1.005-20.10, p = 0.047), increasing age (OR: 1.035; 95% CI 1.022-1.049; p < 0.0001), and shorter hospital stays (OR: 0.7838; 95% CI 0.758-0.811; p < 0.0001). Smoking history (OR: 0.825; 95% CI 0.573-1.187, p > 0.05) and vasospasm (OR: 1.005; 95% CI 0.648-1.558; p > 0.05) were not significantly associated with higher odds of mortality. Mortality following aneurysmal SAH is associated with posterior circulation aneurysms, and increasing age, but not smoking history or vasospasm. These findings may be useful for prognostication and counseling patients and families.


A quantitative use of the NIDCAP tool. The effect of gender and race on very preterm neonates' behavior.
The purpose of this study was to demonstrate how the check sheet of the Newborn Individualized Developmental Care and Assessment Program (NIDCAP) can be quantified and used in research. Using the quantified NIDCAP measures, the hypothesis that Caucasian male infants are less behaviorally competent while in a neonatal intensive care unit (NICU) was explored. Participants included 42 very preterm NICU infants. Eighty-five NIDCAP behaviors were quantified into scores ranging from 0 to 1, indicating the percentage of time each behavior was observed. Multivariate analyses were used in grouping the 85 NIDCAP behaviors into three subsystems of functioning. The hypothesis that Caucasian male infants were less competent was not supported; neither were gender differences found. Contrary to this hypothesis, African American infants were identified as more vulnerable on several behaviors. NIDCAP assessments were easily quantified. Once the entire NIDCAP exam has been quantified, practice applications may involve automated development of individualized care plans.


Organotins in the sediments of the Zuari estuary, west coast of India.
Industrial use of organotins such as butyltins and phenyltins has increased several folds during the last two decades. Butyltins and phenyltins are synthetic, multipurpose chemicals, which have been extensively used in marine antifouling paints. They have been known to be extremely poisonous to mollusc fishery resources (oysters, clams, etc.). Contamination of Zuari estuary sediments was assessed by quantitative determination of butyltins and phenyltins by using GC-MS using the electron ionization mode. Butyltins predominated in the whole area over phenyltins. Butyltins contributed about 70-90% of the organotins in general. The concentration of butyltins in sediments ranged from 20 to 7621 ng Sn/g. The concentration of phenyltins in sediments ranged from 0 to 46 ng Sn/g. Degradation indices for butyltins and phenyltins were calculated. The Butyltin degradation index (BDI) for the Zuari sediments ranged from 0 to 2.7 indicating a lot of fresh input of butyltins in the estuary and a lower degradation rate. The phenyltin degradation index (PhDI) ranged from 1 to 10 implying that different processes were prevalent at different stations. The observed organotin levels in the Zuari estuary indicate some highly localized areas of contamination which are severe enough to cause harmful effects on marine flora and fauna. Therefore, there is a need to regulate the use of butyltins and phenyltins as biocides in marine antifouling paints.


Subareolar injection of 99mTc facilitates sentinel lymph node identification.
Sentinel lymph node (SLN) biopsy with the standard intraparenchymal injection has been accepted as an alternative to routine axillary dissection for patients with breast cancer. However, the identification and false-negative rates with this method can vary widely from surgeon to surgeon. The subareolar location contains a rich lymphatic network and represents a potential site of injection for SLN identification. Between August 1, 1999, and December 31, 2000, we performed 159 SLN biopsy procedures on 158 patients with breast cancer. For each patient, 99mTc-labeled sulfur colloid was injected into the subareolar location, and 1% isosulfan blue dye was given as an intraparenchymal injection. In every case, at least one radioactive SLN was identified with the subareolar injection of technetium; a blue SLN was found in 97% of the cases. The blue SLN was also radioactive in 98% of the cases, indicating that the blue dye injected around the tumor and the technetium injected into the subareolar location drained to the same SLN. Subareolar injection of technetium can improve SLN identification rates for breast cancer. The simplicity and accuracy of this technique may also reduce the variable results reported with the standard intraparenchymal method.


Exploring the prescribing trends and factors affecting initial anti-parkinsonian drug selection in Korea: A nationwide population-based cohort study.
Parkinson's disease (PD) is a common neurodegenerative disorder typically treated with dopamine replacement therapy and dopamine agonists (DAs) to alleviate symptoms and minimize dyskinesia. Optimal treatment strategies for patients newly diagnosed with PD have been a topic of debate for many years. We conducted a 10-year descriptive study of drug prescription trends and factors affecting prescription choices for newly diagnosed drug-naïve PD patients using data from the National Health Insurance program in Korea. To identify statistically significant differences in yearly trends, we employed the Cochran-Armitage trend test. Additionally, we utilized multiple logistic regression analysis to investigate the factors associated with the selection of levodopa and DAs as initial anti-parkinsonian drugs. A total of 99,118 patients with PD who were prescribed levodopa or DAs alone as initial anti-parkinsonian drugs between 2011 and 2020 were eligible for inclusion in the analysis. The prescription rate of DAs increased until 2012, and then steadily decreased annually. The likelihood of levodopa prescription increased with age and at higher-level hospitals. In terms of comorbidities, patients with Alzheimer's disease and cerebrovascular diseases were more likely to be prescribed levodopa than those with peptic ulcer disease and dyslipidemia. The decline in levodopa prescriptions was reversed in 2012, and the prescription rate has continued to increase until recently. The odds ratio of levodopa prescription increased in elderly patients with Alzheimer's disease and decreased in patients with Medical aid insurance and peptic ulcer disease.


Pharmacodynamics, clinical indications, and adverse effects of heparin.
Heparins are a heterogenous group of naturally occurring glycosaminoglycans characterized by anticoagulant activity and a wide range of molecular weights (low molecular weight or fractionated heparins evolving within the past two decades). Cofactors for endogenous inhibitors of coagulation (antithrombin III and heparin cofactor II), heparin administration results in a hypocoagulable state. Various platelet activities, including inhibition of activity induced by platelet-derived growth factors on vascular smooth muscle, also have been noted. Divorced of anticoagulant nature, novel applications may include a role in atherosclerosis prevention, acceleration of collateral coronary as well as peripheral circulation (i.e., angiogenesis), and continued (chronic) post-myocardial infarction therapy. Established indications include treatment of various thrombotic diseases, unstable angina, and thrombosis chemoprophylaxis in medical/surgical patients. The antithrombotic potential of the heparins is used also in thrombosis management related to extracorporeal circulatory assistance or dialysis devices. Heparin's therapeutic potential in the postphlebitic syndrome as well as in acute treatment of myocardial infarction (primarily and adjunctively with various thrombolytic agents) continues to undergo evaluation; however, early data review shows favorable trends for its inclusion in situations that favor thrombus generation (e.g., anterior myocardial infarction). Although associated with thrombocytopenia or hypertransaminasemia, the heparins are relatively well tolerated. In a small subset of patients, a severe thrombocytopenia may ensue, which generally resolves on medication withdrawal. As this class of glycosaminoglycans becomes better characterized, new indications may emerge for both native and the newer fractionated heparins.


An error-prone T7 RNA polymerase mutant generated by directed evolution.
Viruses replicate their genomes at exceptionally high mutation rates. Their offspring evolve rapidly and therefore, are able to evade common immunological and chemical antiviral agents. In parallel, virus genomes cannot tolerate a further increase in mutation rate: Experimental evidence exists that even few additional mutations are sufficient for the extinction of a viral population. A future antiviral strategy might therefore aim at increasing the error-producing capacity of viral replication enzymes. We employed the principles of directed evolution and developed a scheme for the stringent positive selection of error-prone polymerase activity. A mutant T7 RNA polymerase with a nucleotide substitution error rate at least 20-fold greater than that of the wild-type was selected. This enzyme synthesized highly heterogeneous RNA products in vitro or in vivo and also decreased the replication efficiency of wild-type bacteriophage T7 during infection.


Benzo(a)pyrene-caused increased G1-S transition requires the activation of c-Jun through p53-dependent PI-3K/Akt/ERK pathway in human embryo lung fibroblasts.
Benzo(a)pyrene (B(a)P) is a potent lung carcinogen mainly derived from tobacco smoking and environmental contamination, however, the molecular mechanisms by which it accelerates the cell cycle progression and induces the abnormal cell proliferation are still far away from understood. Our current analysis of human embryo lung fibroblasts (HELF) showed that B(a)P exposure was able to promote cell cycle G(1)-S phase transition. This effect was correlated with c-Jun activation because inhibition of c-Jun by its dominant negative mutant (TAM67) reversed B(a)P action on cell cycle with the down-regulation of expression of cyclin D1, pRb and E2F1. Further study found that overexpression of dominant negative mutants of, PI-3K or Akt, dramatically reduced B(a)P-induced the activation of c-Jun and extracellular signaling regulated kinase (ERK), but not c-Jun NH2 terminal kinase (JNK). Inhibition of p53 by either its inhibitor pifithrin-alpha or p53 siRNA markedly increased B(a)P-induced the activation of c-Jun, Akt and ERK in this context. Take together, our results indicate that c-Jun activation by p53-dependent PI-3K/Akt/ERK pathway is responsible for B(a)P-induced cell cycle alternations in human embryo lung fibroblasts.


Robotic Versus Laparoscopic Radical Trachelectomy for Early-Stage Cervical Cancer: Case Report and Review of Literature.
We conducted a literature review to evaluate the minimally invasive fertility-sparing procedures, namely robotic radical trachelectomy (RRT) and laparoscopic radical trachelectomy (LRT), in patients with early-stage cervical cancer. We searched PubMed, MEDLINE, Ovid, Google Scholar, and Scopus up to July 2015 using the following key words and their combinations: cervical cancer, early stage, fertility-sparing surgery, radical trachelectomy, robotic trachelectomy, and laparoscopic trachelectomy. Papers providing details of RRT and LRT separately were included. Extracted papers and their bibliographies were reviewed according to the purpose of the study, and demographic, surgical, and clinical parameters were analyzed. Our review comprised 45 cases of RRT and 216 cases (including our case) of LRT. The median (range) patient age was 29 (9) years in the RRT group and 32 (10) years in the LRT group (p < .001). Histological types and stages were significantly different in the 2 groups (p < .001 for both). The median length of excised parametrial tissue was significantly higher in the RRT group (p < .001). The hysterectomy conversion rate on the results of frozen section examination was significantly higher in the RRT group (37% vs 6.5%; p < .001). There was a significant difference in median lymph node count based on the surgical approach (RRT: 22 [range, 21] vs LRT: 32 [range, 14]; p = .02). Estimated blood loss and length of hospital stay were significantly higher in the LRT group (both p < .001). The mean (range) operative time was 308 (188) minutes in the RRT group and 296 (143) minutes in the LRT group (p < .001). Pregnancy, preterm, and term birth rates were similar in the 2 groups (RRT: 18.5%, 7.4%, and 0, respectively; LRT: 29%, 8%, and 8%, respectively). Thirteen women experienced recurrent cervical cancer in the LRT group, and no recurrence was seen in the RRT group. The median (range) duration of follow-up was 8 (7.5) months in the RRT group and 34 (20) months in the LRT group (p < .001). Based on the reported data, LRT seems comparable to RRT for treating patients with early-stage cervical cancer who wish to preserve fertility.


Prognostic factors in children undergoing salvage surgery for bladder/prostate rhabdomyosarcoma.
Prognostic factors have been studied in patients with rhabdomyosarcoma in general, but little is known about prognostic factors in the subgroup of patients with bladder/prostate rhabdomyosarcoma (BP-RMS) requiring salvage surgery after failure of chemotherapy ± radiotherapy to achieve local control. We reviewed the 28-year Italian experience with BP-RMS requiring salvage surgery after failure of nonsurgical management indicated by evidence of disease persistence after chemotherapy ± radiotherapy. Our hypothesis was that the same variables identified as prognostic factors in the general population with RMS could have prognostic value in this subgroup. Between 1986 and 2014, 108 patients with a histological diagnosis of BP-RMS were registered into three consecutive protocols. Patients undergoing salvage surgery after failure of chemotherapy ± radiotherapy and follow-up >5 years were considered for study. Variables related to the patient, to treatment, and to the surgical specimen were compared using the log-rank test in patients who achieved and failed to achieve 5-year progression-free survival (5-yr PFS). Odds ratios (OR) and 95% confidence intervals (95% CI) were calculated for significant variables. Of the 108 patients enrolled in the protocols, 33 (30.6%) underwent salvage surgery and 26 matched study criteria. Seventeen patients were disease-free after a median follow-up of 125 months (range 65.5-311.7 months), 5-yr PFS 65.4%. Nine events were registered after a median follow-up of 5.4 months (range 1.9-27.5 months). Among the variables assessed (Table), only an undifferentiated histology proved to be significantly associated with a poorer 5-yr PFS, whereas a tumor size above 5 cm in the removed specimen approached significance. The OR (95% CI) for failure of 5-yr PFS was 28 (2.4-326.8) and 8.3 (1.03-67.1), respectively. 5-yr PFS proved unrelated to excision margins of the surgical specimen, that is on whether there was evidence of microscopic residues left behind during surgery. These data suggest that the outcome is more influenced by the biological properties of the tumor. If small foci of differentiated cells are left behind, these probably do not compromise final prognosis. In our patients requiring salvage bladder-prostate surgery after failure of chemotherapy ± radiotherapy, long-term progression-free survival seemed unrelated to patient/tumor characteristics at presentation and preoperative management. Regarding the characteristics of the removed specimen, instead, an undifferentiated tumor histology and a diameter of the removed tumor >5 cm negatively influenced prognosis, whereas the presence of positive excision margins did not.


Imaging neuronal structure dynamics using 2-photon super-resolution patterned excitation reconstruction microscopy.
Visualizing fine neuronal structures deep inside strongly light-scattering brain tissue remains a challenge in neuroscience. Recent nanoscopy techniques have reached the necessary resolution but often suffer from limited imaging depth, long imaging time or high light fluence requirements. Here, we present two-photon super-resolution patterned excitation reconstruction (2P-SuPER) microscopy for 3-dimensional imaging of dendritic spine dynamics at a maximum demonstrated imaging depth of 130 μm in living brain tissue with approximately 100 nm spatial resolution. We confirmed 2P-SuPER resolution using fluorescence nanoparticle and quantum dot phantoms and imaged spiny neurons in acute brain slices. We induced hippocampal plasticity and showed that 2P-SuPER can resolve increases in dendritic spine head sizes on CA1 pyramidal neurons following theta-burst stimulation of Schaffer collateral axons. 2P-SuPER further revealed nanoscopic increases in dendritic spine neck widths, a feature of synaptic plasticity that has not been thoroughly investigated due to the combined limit of resolution and penetration depth in existing imaging technologies.


The pharmacist's role in chimeric antigen receptor T cell therapy.
The emergence and efficacy of chimeric antigen receptor (CAR) T cell therapy in previously incurable malignancies represents a promising paradigm shift in cancer care. However, it is not without significant clinical, operational, and financial considerations. Pharmacists should be prepared to fulfill the various roles in CAR T cell therapy provision including: policy development; electronic medical record build; patient and staff education; patient selection; procurement, storage, and handling; medication administration and supportive care; management of adverse reactions; and quality tracking. Our commentary provides an overview of the opportunities for pharmacy involvement in the implementation and maintenance of a CAR T cell therapy program with an emphasis on the importance of pharmacy involvement as part of a multidisciplinary approach to care. Although some institutions have dedicated a CAR T cell pharmacist to meet the demands of emerging CAR T cell therapy, we believe that clinical pharmacists practicing in hematopoietic stem cell transplant and hematology/oncology have the skills and training to fulfill the pharmacist's role in CAR T cell therapy.


Outbreak of bacterial meningitis among patients undergoing myelography at an outpatient radiology clinic.
To investigate an outbreak of bacterial meningitis at an outpatient radiology clinic (clinic A) and to determine the source and implement measures to prevent additional infections. A case was defined as bacterial meningitis in a patient undergoing myelography at clinic A from October 11 to 25, 2010. Patients who underwent myelography and other procedures at clinic A during that period were interviewed, medical records were reviewed, and infection prevention practices were assessed. Case-patient cerebrospinal fluid (CSF) specimens, oral specimens from health care personnel (HCP), and opened iohexol vials were tested for bacteria. Bacterial isolates were compared using pulsed-field gel electrophoresis. A culture-negative CSF specimen was tested using a real-time polymerase chain reaction assay. Three cases were identified among 35 clinic A patients who underwent procedures from October 11 to 25, 2010. All case-patients required hospitalization, 2 in an intensive care unit. Case-patients had myelography performed by the same radiology physician assistant and technician on October 25; all patients who underwent myelography on October 25 were affected. HCP did not wear facemasks and reused single-dose iohexol vials for multiple patients. Streptococcus salivarius (a bacteria commonly found in oral flora) was detected in the CSF of 2 case-patients (1 by culture, 1 using real-time polymerase chain reaction) and in HCP oral specimens; 1 opened iohexol vial contained Staphylococcus epidermidis. Pulsed-field gel electrophoresis profiles from the case-patient S salivarius and the radiology physician assistant were indistinguishable. Bacterial meningitis likely occurred because HCP performing myelography did not wear facemasks; lapses in injection practices may have contributed to transmission. Targeted education regarding mask use and safe injection practices is needed among radiology HCP.


A novel trophectoderm biopsy technique for all blastocyst stages.
This study was conducted to assess the effectiveness of a new trophectoderm (TE) biopsy method that does not require prior opening of the zona pellucida at the blastocyst stage. TE biopsy was conducted using a modified extrusion method for embryos during the cleavage stage. In this method, culture medium was injected into the perivitelline space to help extrude TE cells from the zona pellucida before TE biopsy. Our extrusion method preserves the embryo culture environment until immediately before biopsy because it does not require opening of the zona pellucida prior to TE biopsy. Furthermore, this method does not require a waiting time for blastocyst hatching after laser irradiation, thereby minimizing damage to the embryos and maintaining the time schedule of culture operations. TE biopsy using this novel extrusion method may be useful in various applications, including the collection of TE cells for next-generation sequencing analysis.


Rib spur causing a hemothorax, pneumothorax, and diaphragmatic injury in a pediatric patient.
Pleural effusion is a relatively common condition encountered in the pediatric emergency department. Evaluation of pleural effusion in the emergency department typically includes advanced imaging such as computer tomography or ultrasound, as well as diagnostic thoracocentesis. We report a case of a 10-year-old female with a rib spur at the anterolateral left sixth rib that caused a hemothorax, pneumothorax, and diaphragmatic injury. The patient underwent video-assisted thoracoscopic surgery and resection of the rib spur. The procedure was well-tolerated without any complications.


Efferent projections from the flocculus in the albino rat as revealed by an autoradiographic orthograde tracing method.
The terminal sites of floccular efferent fibers were investigated in the albino rat by an autoradiographic orthograde method. The corticonuclear fibers terminated in the caudoventral part of the lateral cerebellar nucleus and in the caudoventral region of the lateral part of the posterior interpositus nucleus. A few fibers from the rostral flocculus terminated in the granular cell layer of the basolateral part of the nodulus and uvula as mossy fiber type terminals. The projection to the nodulus and uvula was confirmed, by an additional retrograde HRP study, to originate from scattered spindle-shaped cells in the floccular stalk. The corticovestibular fibers terminated massively in the subnucleus y. The fibers passing through the subnucleus y divided into two bundles; one bundle coursed rostrally to terminate in the lateral and ventral parts of the superior vestibular nucleus, while the other bundle passed through the lateral and then ventral parts of the lateral vestibular nucleus, supplying a few terminals to these regions, to terminate sparsely in the rostral to intermediate part of the medial vestibular nucleus and the rostroventral part of the spinal vestibular nucleus. Some fibers passing through the lateral vestibular nucleus coursed rostrally to terminate in the medial part of the superior vestibular nucleus. Sparse terminals derived from the rostral flocculus were found in the prepositus hypoglossal nucleus. No definitive differential efferent projections were demonstrated in the rat flocculus.


Hydro-organic mobile phase and factorial design application to attain green HPLC method for simultaneous assay of paracetamol and dantrolene sodium in combined capsules.
The greenness of any analytical method has become a very important aspect of a good analytical method. However, most chromatographic methods depend on the usage of relatively large amounts of lethal and un-decaying chemicals and solvents. So, a green approach based on the full factorial design was employed to develop a simple and rapid HPLC technique for concurrent determination of paracetamol and dantrolene sodium in their combined capsules. Both drugs are highly recommended to be administered together in patients with severe musculoskeletal disorders. Avoiding the routine methodology and resorting to the modern technology represented in the usage of experimental design allows rapid determination of the studied drugs using the optimum quantity of chemicals to avoid any waste of resources. Simultaneous separation of a binary mixture of paracetamol and dantrolene sodium was accomplished using a reversed phase Hypersil C18 column using an eco-friendly isocratic eluent. The used mobile phase consisted simply of ethanol: water (40:60, v/v). Orthophosphoric acid was used to adjust the pH of the mobile phase to 4.5. Triethanolamine (0.2%) was added aiming to reduce the peak tailing. The assay was completed within less than 6 min adopting 0.8 mL/min as a flow rate. The detection was carried out using a UV-detector at 290 nm. The suggested technique shows a linear correlation over concentration ranges of 1.0-200 and 1.0-40 µg/mL for paracetamol and dantrolene sodium, respectively. The suggested technique allowed the simultaneous analysis of the two co-formulated drugs in their synthetic mixture and combined capsule. The suggested technique is considered a greener substitute for the other reported HPLC techniques through the usage of safer solvents and chemicals, along with decreasing both waste output and analysis time. The method is accurate with recoveries between 97.85 and 101.27%, precise, as %RSD for the intraday and interday precision were between 0.39 and 1.72% and very sensitive with limits of detection (LOD)'s 0.15 and 0.18 µg/ml and limits of quantification (LOQ)'s 0.48 and 0.61 µg/ml for paracetamol and dantrolene sodium, respectively. The method greenness was ensured through its assessment by four greenness metrics. It is also validated following the International Conference on Harmonization Guidelines. The recommended technique could be a good alternative to traditional methods in the routine quality control analysis of the studied drugs due to its minimum harm to the planet or human beings.


Haematological manifestations of arthrogryposis-renal dysfunction-cholestasis (ARC) syndrome: a case report.
Arthrogryposis multiplex congenita, renal dysfunction, and cholestasis (ARC) syndrome is a rare, fatal, multisystem disorder. Bleeding problems, which occur spontaneously and post organ biopsies, have been reported in these patients. We report the case of an infant who had life-threatening spontaneous nasal bleeding. A detailed assessment of her platelet function and morphology is presented.


FGF-1 delivery from multilayer alginate microbeads stimulates a rapid and persistent increase in vascular density.
In recent years, great advances have been made in the use of islet transplantation as a treatment for type I diabetes. Indeed, it is possible that stimulation of local neovascularization upon transplantation could improve functional graft outcomes. In the present study, we investigate the use of multilayered alginate microbeads to provide a sustained delivery of FGF-1, and whether this results in increased neovascularization in vivo. Multilayered alginate microbeads, loaded with either 150ng or 600ng of FGF-1 in the outer layer, were surgically implanted into rats using an omentum pouch model and compared to empty microbead implants. Rats were sacrificed at 4days, 1week, and 6weeks. Staining for CD31 showed that both conditions of FGF-1 loaded microbeads resulted in a significantly higher vessel density at all time points studied. Moreover, at 6weeks, alginate microbeads containing 600ng FGF-1 provided a greater vascular density compared to both the control group and the microbeads loaded with 150ng FGF-1. Omenta analyzed via staining for smooth muscle alpha actin showed no variation in mural cell density at either 4days or 1week. At 6weeks, however, omenta exposed to microbeads loaded with 600ng FGF-1 showed an increase in mural cell staining compared to controls. These results suggest that the sustained delivery of FGF-1 from multilayered alginate microbeads results in a rapid and persistent vascular response. An increase in the local blood supply could reduce the number of islets required for transplantation in order to achieve clinical efficacy.


Six complementation groups for ionising-radiation sensitivity in Chinese hamster cells.
The ionising radiation-sensitive mutants irs 1, irs 2, irs 3, xrs-1 (or xrs-7), EM7 and XR-1 were fused to wild-type cells or to each other in pairs to create hybrid cells. These hybrids were checked chromosomally and their X-ray sensitivity tested. Each mutant was found to be recessive to wild-type and to complement the X-ray sensitivity of the other mutants. Thus there appear to be at least 6 complementation groups for ionising radiation sensitivity in Chinese hamster cells.


ST2 is an inhibitor of interleukin 1 receptor and Toll-like receptor 4 signaling and maintains endotoxin tolerance.
The Toll-interleukin 1 receptor (TIR) superfamily, defined by the presence of an intracellular TIR domain, initiates innate immunity through activation of the transcription factor NF-kappa B, leading to the production of proinflammatory cytokines. ST2 is a member of the TIR family that does not activate NF-kappa B and has been suggested as an important effector molecule of T helper type 2 (T(H)2) responses. We show here that the membrane-bound form of ST2 negatively regulated type I interleukin 1 receptor (IL-1RI) and Toll-like receptor 4 (TLR4) but not TLR3 signaling by sequestrating the adaptors MyD88 and Mal. In contrast to wild-type mice, ST2-deficient mice failed to develop endotoxin tolerance. Thus, these results provide a molecular explanation for the function of ST2 in T(H)2 responses, as inhibition of TLRs promotes a T(H)2 response, and also identify ST2 as a key regulator of endotoxin tolerance.


Electronic medical records as a tool in clinical pharmacology: opportunities and challenges.
The development and increasing sophistication of electronic medical record (EMR) systems hold the promise of not only improving patient care but also providing unprecedented opportunities for discovery in the fields of basic, translational, and implementation sciences. Clinical pharmacology research in the EMR environment has only recently started to become a reality, with EMRs becoming increasingly populated, methods to mine drug response and other phenotypes becoming more sophisticated, and links being established with DNA repositories.


Cognitive impairment affects physical recovery of patients with heart failure undergoing intensive cardiac rehabilitation.
Purpose. To determine whether the presence of cognitive impairment (CI) affects physical recovery of patients with chronic heart failure (CHF) undergoing a cardiac rehabilitation program (CRP). Methods. We enrolled 80 CHF patients (M/F = 53/27). CI was evaluated by means of the Mini-Mental State Examination (MMSE), exercise tolerance was evaluated by six-minute walking test (6 mwt). All patients underwent a 6-week CRP program at 50-70% of maximal V(O2). Patients were divided into two groups according to their MMSE (group 1: 16-23; group 2: 24-30). Results. MMSE resulted directly related to ejection fraction (r = 0.42; P = 0.03), and it was inversely related to creatinine (r = -0.36; P = 0.04). At 6 week group 1 had a lower increase in distance walked at 6 MWT than group 2 (P = 0.008). At multivariate logistic regression MMSE 16-23 predicted a reduced exercise recovery in the overall population (OR = 1.84; 95% CI = 1.50-2.18) and in women (OR = 1.42; 95% CI = 1.22-1.75), while it was not predicted in males. Conclusions. CI is a marker of advanced CHF and is an independent predictor of lower exercise recovery after CRP.


Micro-computerized tomographic analysis of premolars restored with oval and circular posts.
The purpose of the study was to evaluate by micro-computerized tomography (microCT) areas and volumes of post, cement, and voids/bubbles in the post space of oval-shaped premolars restored either with oval or circular posts. Twelve extracted premolars were divided into two groups according to the drill-fiber post system used: (1) GC Fiber Post Drill + circular post GC Fiber Post; (2) Ellipson tip(TM) + oval post Ellipson post(TM). Each tooth was scanned using microCT, and areas and volumes of canal, post space, post, cement, and voids at coronal, medium, and apical level were calculated by using a three-dimensional visualization software. Two-way analyses of variance and Tukey tests were used for statistical analysis (p < 0.05). The area of voids was significantly greater at apical level of oval posts, but no difference was found between the levels among the groups. Regardless of post shape, the volume of voids and of cement was significantly higher at the coronal level. With oval posts, the total volume of cement was twice as much as with circular posts, and the difference was statistically significant. Our results indicate that the volume of voids does not depend on post shape. Moreover, the microCT analysis demonstrated that the volume of cement was greater with oval posts compared to circular posts. The microCT analysis provided interesting information on voids distribution and on the ratio between post shape and cement thickness. These results might address the clinician in the reconstruction of teeth with fiber posts.


IGF-1-containing multi-layered collagen-fibrin hybrid scaffolds for bladder tissue engineering.
Clinical success of bladder reconstructive procedures could be promoted by the availability of functional biomaterials. In this study, we have developed a multi-layered scaffold consisting of a bioactive fibrin layer laminated between two collagen sheets all having undergone plastic compression. With this construct we performed bladder augmentation in a nude rat model after partial bladder excision and evaluated the morphological and functional behavior of the implant. The fibrin was functionalized with a recombinant human insulin-like growth factor-1 (IGF-1) variant that covalently binds fibrin during polymerization and has a matrix metalloproteinase-cleavage insert to enable cell-mediated release. The purified IGF-1 variant showed similar bioactivity in vitro compared to commercially available wild type (wt) IGF-1, inducing receptor phosphorylation and induction of human smooth muscle cell proliferation. In vivo, the multi-layered bioactive collagen-fibrin scaffolds loaded with the IGF-1 variant triggered dose-dependent functional host smooth muscle cell invasion and bundle formation with re-urothelialization 4weeks after surgery in a rat model. The design of new bio-functional scaffolds that can be employed for bladder reconstructive procedures is a growing focus in the field of tissue engineering. In this study, a fibrin binding form of human insulin-like growth factor-1 (IGF-1) was produced and used to functionalize a multi-layered collagen-fibrin scaffold consisting of bioactive fibrin layer, sandwiched between two collagen gels. An effective dosage of our IGF-1 variant was successfully determined via a nude rat bladder model, which may play a critical role in estimating its therapeutic dosage in clinical trials. Thus, this new bioactive scaffold may offer an advanced approach to accelerate bladder regeneration.


Photoinduced electron transfer from N,N-dimethylaniline to 7-amino coumarins in protein-surfactant complex: slowing down of electron transfer dynamics compared to micelles.
Photoinduced electron transfer from N,N-dimethylaniline to different Coumarin dyes has been investigated in dodecyl trimethyl ammonium bromide (DTAB) micelles and in Bovine serum albumin (BSA)-DTAB protein-surfactant complex using steady-state and picosecond time-resolved fluorescence spectroscopy. We observed a slower fluorescence quenching rate in the DTAB micelles and in the protein-surfactant complex as compared to that in pure acetonitrile solution. Moreover, the observed fluorescence quenching in BSA-DTAB complex was found to be slower than that in DTAB micelles. In the correlation of free-energy change with the fluorescence quenching constant we observed a deviation in the fluorescence quenching electron transfer rate for Coumarin 151 (C-151) from the normal Marcus curve. This observation is ascribed to the stronger interaction of C-151 with the surfactant molecules present in the micelles. This is evident from the slower translation diffusion (D(L)) of Coumarin 151 compared to other probe molecules.


Off-Label 9-Valent Human Papillomavirus Vaccination for Actinic Keratosis: A Case Series.
The suspected link between human papillomavirus (HPV) and the development of premalignant and malignant skin lesions remains inadequately examined in clinical settings. This case series describes HPV vaccination as an off-label adjuvant therapy for actinic keratosis (AK). Twelve immunocompetent AK patients underwent HPV vaccination at a private dermatology clinic in Naestved, Denmark. Prior to vaccination, all patients demonstrated a high AK burden that required regular control visits. At 0, 2, and 6 months, the patients received an intramuscular injection of a commercially available 9-valent HPV vaccine. Concurrently, patients continued conventional AK therapies at 3-month intervals. Clinical response, consisting of reduction in AK number and general change in skin appearance, was assessed by a dermatologist over 12 months following first vaccination. All patients (mean age 76.2 years; 10 M and 2 F) completed the vaccine schedule. Overall, an average 85% reduction in total AK burden was recorded 12 months after beginning vaccination. Median AK burden thus fell from 56 (IQR: 44-80) to 13.5 (IQR: 1-18) lesions after 12 months. Lesion reduction was observable by the second inoculation at month 2 (34 AKs; IQR 22-80), continuing steadily until month 6 (15 AKs; IQR 5-30) and plateauing between 6 and 12 months. Clinically, HPV vaccination elicited fading of lesions' erythematous background after the first dose, often followed by sloughing of hyperkeratotic elements in subsequent weeks. Patients reported no adverse effects related to HPV vaccination. This case series introduces the possibility that 9-valent HPV vaccination in combination with conventional treatments may be used as a therapeutic strategy for AK.


Granulocyte colony stimulating factor treatment for delayed recovery of ticlopidine-related neutropenia.
We describe the case of a patient who developed neutropenia associated with sepsis and endophthalmitis after ticlopidine therapy for coronary stenting. The neutropenia did not resolve until granulocyte colony stimulating factor (G-CSF) was given. This uncommon case brings to attention the need for the immediate use of G-CSF in patients with delayed recovery from drug-related neutropenia and severe infection.


Amyloid β: Walking on the dark side of the moon.
For some decades, amyloid β (Aβ) has only been considered as a cytotoxic peptide, putative cause and marker of Alzheimer's disease (AD). Today, however, a considerable amount of evidence goes against the classical amyloid hypothesis and illustrates a new picture in which the Aβ loss of function, rather than its accumulation, has a pathogenic role in AD. In this concise review, we summarize some highlights of a collection of research pointing to the physiological function of Aβ and its role in the mechanisms of memory formation.


Spectrum of scalar and pseudoscalar glueballs from functional methods.
We provide results for the spectrum of scalar and pseudoscalar glueballs in pure Yang-Mills theory using a parameter-free fully self-contained truncation of Dyson-Schwinger and Bethe-Salpeter equations. The only input, the scale, is fixed by comparison with lattice calculations. We obtain ground state masses of 1.9 GeV and 2.6 GeV for the scalar and pseudoscalar glueballs, respectively, and 2.6 GeV and 3.9 GeV for the corresponding first excited states. This is in very good quantitative agreement with available lattice results. Furthermore, we predict masses for the second excited states at 3.7 GeV and 4.3 GeV . The quality of the results hinges crucially on the self-consistency of the employed input. The masses are independent of a specific choice for the infrared behavior of the ghost propagator providing further evidence that this only reflects a nonperturbative gauge completion.


Intuitive eating Scale-2: Factor structure and associations with disordered eating, impulsivity and quality of life in adolescents with overweight/obesity.
The Intuitive Eating Scale-2 (IES-2) has been shown to be a valid tool to assess the capability of eating in reaction to natural hunger/satiety cues. However, its factor structure seems to differ in function of cultural/socioeconomic backgrounds, and its psychometric properties among the adolescents with overweight/obesity (BMI-for-age percentile >85th) have not been examined yet. Thus, this study aims to 1) investigating the factorial structure/psychometric properties of IES-2 in adolescents with overweight/obesity; and 2) exploring the associations between impulsivity, quality of life disordered and intuitive eating. A total of 202 Portuguese adolescents (124 girls; 78 boys; 12-19 years) under weight-loss treatment with a mean BMI z-score of 2.41 (SD = 0.75) participated in this study. The IES-2 factor structure was explored by confirmatory factor analysis and bifactor models. Test-retest reliability analyses were performed over 6 months (n = 41) and associations between the variables under study were explored. Confirmatory factor analyses with posthoc modifications resulted in a bifactor model with acceptable fit supporting one general factor (intuitive eating) and three specific factors (IES-2 subscales). The "Unconditional Permission to Eat" subscale could not be replicated in this sample. Test-retest reliability analyses suggested good temporal stability. Intuitive eating scores were negatively associated with grazing eating behavior and impulsivity (negative urgency) and positively linked to quality of life. An adjusted version of IES-2 can be an appropriate measure for assessing intuitive eating levels in adolescents with overweight/obesity. Research on intuitive eating has the potential to enhance pediatric weight-loss interventions.


Electronic tuning of Mg2Cu6Ga5. A route to crystalline approximant and quasicrystalline phases.
Studies of Mg2Cu6Ga5 reveal that this compound contains incomplete Bergman clusters in its structure and shows a pseudogap and empty bonding states just above the Fermi energy according to band calculations. Under a rigid band assumption, such a compound may be tuned to approximant and quasicrystal phases in which the required number of electrons are attained. Here, we replace part of Mg in the isotypic Mg2Cu6Ga5 with Sc, and both 1/1 approximant and icosahedral quasicrystal phases are obtained after some fine-tuning. This method closely correlates the pseudogap and bonding with Hume-Rothery concepts, thus giving useful directions for future quasicrystal searches, especially when approximants are not known.


Prospective randomized trial of preoperative enteral immunonutrition followed by elective total gastrectomy for gastric cancer.
Perioperative enteral immunonutrition is thought to reduce postoperative morbidity in patients undergoing major gastrointestinal surgery. This study assessed the clinical effects of preoperative enteral immunonutrition in well nourished patients with gastric cancer undergoing total gastrectomy. Well nourished patients with primary gastric cancer, fit for total gastrectomy, were randomized to either a control group with regular diet, or an immunonutrition group that received regular diet supplemented with 1000 ml/day of immunonutrients for 5 consecutive days before surgery. The primary endpoint was the incidence of surgical-site infection (SSI). Secondary endpoints were rates of infectious complications, overall postoperative morbidity and C-reactive protein (CRP) levels on 3-4 days after surgery. Of 244 randomized patients, 117 were allocated to the control group and 127 received immunonutrition. SSIs occurred in 27 patients in the immunonutrition group and 23 patients in the control group (risk ratio (RR) 1.09, 95 per cent confidence interval 0.66 to 1.78). Infectious complications were observed in 30 patients in the immunonutrition group and 27 in the control group (RR 1.11, 0.59 to 2.08). The overall postoperative morbidity rate was 30.8 and 26.1 per cent respectively (RR 1.18, 0.78 to 1.78). The median CRP value was 11.8 mg/dl in the immunonutrition group and 9.2 mg/dl in the control group (P = 0.113). Five-day preoperative enteral immunonutrition failed to demonstrate any clear advantage in terms of early clinical outcomes or modification of the systemic acute-phase response in well nourished patients with gastric cancer undergoing elective total gastrectomy. ID 000000648 (University Hospital Medical Information Network (UMIN) database).


DNA microarray analysis of the heat shock transcriptome of the obligate intracytoplasmic pathogen Rickettsia prowazekii.
Here we present the first oligonucleotide DNA microarray analysis of global gene expression changes in the obligate intracytoplasmic pathogen Rickettsia prowazekii using temperature upshift as a model stress condition, and we describe a methodology for isolating highly purified rickettsial RNA. In toto, 23 transcripts were significantly increased by temperature upshift (> or = 2.0-fold; P < 0.05), and no transcripts demonstrated reproducible decreases. Array results for three heat shock-inducible mRNAs were confirmed using quantitative reverse transcription-PCR.


An electron microscopic study of the small cutaneous vessels in lepromatous leprosy.
The small dermal vessels play an important role for the propagation of Mycobacterium leprae. Bacteria can multiply in the endothelial cells as well as in the pericytes and can enter the blood stream from the endothelial cells and the interstitial space from the pericytes. M. leprae can be phagocytosed by blood monocytes.


Paired opposite clear corneal incisions to correct preexisting astigmatism in cataract patients.
To evaluate the astigmatic correcting effect of paired opposite clear corneal incisions on steep axis in cataract patients. Sligo General Hospital, Sligo, Ireland. Fifteen eyes of 14 cataract patients with a mean age of 78.4 years +/- 6.38 (SD) (range 69 to 90 years) were recruited for the study. Inclusion criterion was topographic astigmatism of more than 2 diopters (D) in the cataractous eye. Preoperative refraction, autokeratometry, and topography were performed. The steep axis was marked before sub-Tenon's anesthesia was given. Paired 3-step self-sealing opposite clear corneal incisions were made 1 mm anterior to limbus on the steep axis with a 3.2 mm keratome. One incision was used for standard phacoemulsification, and the other was left unused for astigmatic correction. All the patients had day-case surgery. The first follow-up was at 1 month. Postoperative topography, keratometry, and refraction were performed on all patients. Mean preoperative and postoperative topographic corneal astigmatism were 3.26 +/- 1.03 D (range 2.30 to 5.80 D) and 2.02 +/- 1.04 D (range 0.20 to 4.00 D), respectively. Mean astigmatic correction was 1.23 +/- 0.49 D (range 0.30 to 2.20 D). Mean surgically induced astigmatism by vector analysis was 2.10 +/- 0.79 D (range 0.80 to 3.36 D). There were no incision-related complications. Paired opposite clear corneal incisions on the steep axis is a useful way to correct astigmatism in cataract patients, requiring no extra skill or instrumentation.


Effect of Human Central Nervous System Stem Cell Subretinal Transplantation on Progression of Geographic Atrophy Secondary to Nonneovascular Age-Related Macular Degeneration.
To evaluate the effect of subretinally transplanted human central nervous system stem cells (HuCNS-SC) on the progression of geographic atrophy (GA) in patients with nonneovascular age-related macular degeneration (AMD). Multicenter, prospective, phase 1 open-label clinical trial. Fifteen patients with bilateral GA solely the result of AMD. The eye with the worst best-corrected visual acuity from each patient was selected for treatment and was considered the study eye; fellow eyes served as controls. A total of 0.25 × 106 or 1.0 × 106 HuCNS-SCs were infused directly into the subretinal space, superotemporal to the fovea near the junctional zone, outside the area of GA. All patients underwent spectral-domain OCT and fundus autofluorescence imaging using the Spectralis HRA+OCT (Heidelberg Engineering, Inc., Heidelberg, Germany). Total GA area in both eyes was measured at baseline and month 12 by certified reading center graders using the Spectralis Region Finder software. Sectoral (clock hour) per directional radial GA progression rates with respect to the foveal center in both eyes were calculated using the polar transformation method in Image J software (National Institutes of Health, Bethesda, MD). To facilitate comparative analysis across the cohort, all eyes were transformed to a right-eye orientation. Total GA area and sectoral per directional GA progression rates were compared in both study and control eyes. No statistically significant difference was found in mean change in total GA area at month 12 between study and fellow eyes (1.07 ± 0.84 mm2 vs. 2.08 ± 1.97 mm2; P = 0.08). However, the month 12 sectoral per directional radial GA growth rate for the superotemporal region (i.e., the location of HuCNS-SC transplantation) showed a significantly slower progression rate in study eyes than in fellow eyes (0.29 ± 0.58 mm vs. 1.08 ± 0.65 mm; P = 0.007). The progression rate in the superotemporal quadrant of the study eye was significantly slower than in the other 3 quadrants combined (P = 0.04). In this small pilot study, HuCNS-SC transplantation seems to be associated with slower expansion of the GA lesion in the transplanted quadrant. Larger confirmatory studies are required. Sectoral or directional analysis of growth rates of GA may be a useful approach for assessing the efficacy of locally delivered therapies.


Patterned poly(lactic acid) films support growth and spontaneous multilineage gene expression of adipose-derived stem cells.
Conventional culture surfaces do not provide optimal environmental cues for expansion or differentiation of adult stem cells. Aiming to increase the efficiency of the in vitro culture conditions, biocompatible and biodegradable biomaterials such as poly(lactic acid) (PLA) have been proposed to engineer the stem cell microenvironment. In this study, we explored the feasibility of using PLA substrates to control the responses of adipose-derived stem cells (ASCs). The substrates consisted of flat and patterned PLA films fabricated by casting a chloroform-PLA solution on a glass surface. Patterning was achieved through the condensation of nano-sized water droplets during chloroform evaporation, which resulted in films displaying irregularly distributed circular indentations with a mean diameter of 248±65 nm. Both types of PLA substrates were assessed for protein adsorption using fibronectin and in vitro cell culturing. Tissue-culture polystyrene (TCPS) plates were used as control surfaces. The experiments demonstrated that the patterned PLA substrates had a significantly higher fibronectin adsorption capacity when compared with the flat counterparts. For the entire duration of the culture period, there was no significant difference in cell growth rate on the PLA surfaces with respect to TCPS despite signs of reduced adhesion. In addition, the semi-quantitative real-time RT-PCR analysis of a set of 14 lineage-specific genes revealed that the PLA-related transcriptional activity significantly surpassed that of TCPS. Remarkably, when assessing the effect of patterning, the patterned films proved superior regarding the activation of genes involved in the skeletal myogenic, cardiomyogenic, chondrogenic, and adipogenic pathways. Taken together, our data provide evidence that the surface patterning can exert such an influence on the stem cell microenvironment that the differentiation process can be effectively modulated. Consequently, the patterned PLA surfaces could potentially be used as a platform for localized delivery and engraftment of stem cells.


Integrity of XIAP is essential for effective activity recovery of apoptosome and its downstream caspases by Smac/Diablo.
Contribution of individual BIR domains to Smac antagonism is investigated. Ammonium citrate was used to activate caspase-9 and pro-caspase-9 (D315, D330/A). However, the presence of citrate resulted in autoproteolysis of pro-caspase-9 and its inhibition by XIAP BIR3, which was not observed for apoptosome activated pro-caspase-9 indicating abnormal behavior of pro-caspase-9 in kosmotropic citrate salt. Thus, we used Apaf-1(residues 1-591) to activate caspase-9 through the formation of mini-apoptosome instead. Inhibition of apoptosome by XIAP BIR-1-2-3 was observed to be similar to that of BIR3 indicating that the cleavage of XIAP does not affect its potency. However, BIR1-2-3 was more prone to Smac antagonism due to simultaneous interaction of two BIR domains from XIAP with two N-terminal binding sites of Smac. Therefore, despite the role in caspase-9 activation, Apaf-1 does not influence caspase-9 inhibition by XIAP. In addition, caspase-3, -7 and -9 activity recovery by Smac protein and peptide were more efficient for BIR1-2-3 than for BIR1-2. Consequently, it can be proposed that the presence of multiple BIR domains for XIAP among different species along with dimeric nature of Smac are evolutionary designed to strengthen the antagonistic activity of Smac culminating in efficient induction of cell death.


Elucidating the cellular uptake mechanisms of heptamethine cyanine dye analogues for their use as an anticancer drug-carrier molecule for the treatment of glioblastoma.
The development of chemotherapies for glioblastoma is hindered by their limited bioavailability and toxicity on normal brain function. To overcome these limitations, we investigated the structure-dependent activity of heptamethine cyanine dyes (HMCD), a group of tumour-specific and BBB permeable near-infrared fluorescent dyes, in both commercial (U87MG) and patient-derived GBM cell lines. HMCD analogues with strongly ionisable sulphonic acid groups were not taken up by patient-derived GBM cells, but were taken up by the U87MG cell line. HMCD uptake relies on a combination of transporter uptake through organic anion-transporting polypeptides (OATPs) and endocytosis into GBM cells. The uptake of HMCDs was not affected by p-glycoprotein efflux in GBM cells. Finally, we demonstrate structure-dependent cytotoxic activity at high concentrations (EC50 : 1-100 μM), likely due to mitochondrial damage-induced apoptosis. An in vivo orthotopic glioblastoma model highlights tumour-specific accumulation of our lead HMCD, MHI-148, for up to 7 days following a single intraperitoneal injection. These studies suggest that strongly ionisable groups like sulphonic acids hamper the cellular uptake of HMCDs in patient-derived GBM cell lines, highlighting cell line-specific differences in HMCD uptake. We envisage these findings will help in the design and structural modifications of HMCDs for drug-delivery applications for glioblastoma.


Slp-76 is a critical determinant of NK-cell mediated recognition of missing-self targets.
Absence of MHC class I expression is an important mechanism by which NK cells recognize a variety of target cells, yet the pathways underlying "missing-self" recognition, including the involvement of activating receptors, remain poorly understood. Using ethyl-N-nitrosourea mutagenesis in mice, we identified a germline mutant, designated Ace, with a marked defect in NK cell mediated recognition and elimination of "missing-self" targets. The causative mutation was linked to chromosome 11 and identified as a missense mutation (Thr428Ile) in the SH2 domain of Slp-76-a critical adapter molecule downstream of ITAM-containing surface receptors. The Slp-76 Ace mutation behaved as a hypomorphic allele-while no major defects were observed in conventional T-cell development/function, a marked defect in NK cell mediated elimination of β2-microglobulin (β2M) deficient target cells was observed. Further studies revealed Slp-76 to control NK-cell receptor expression and maturation; however, activation of Slp-76(ace/ace) NK cells through ITAM-containing NK-cell receptors or allogeneic/tumor target cells appeared largely unaffected. Imagestream analysis of the NK-β2M(-/-) target cell synapse revealed a specific defect in actin recruitment to the conjugate synapse in Slp-76(ace/ace) NK cells. Overall these studies establish Slp-76 as a critical determinant of NK-cell development and NK cell mediated elimination of missing-self target cells in mice.


MiR-125b but not miR-125a is upregulated and exhibits a trend to correlate with enhanced disease severity, inflammation, and increased mortality in sepsis patients.
This study aimed to investigate the correlation of miR-125a/b expressions with disease risk, progression, and prognosis of sepsis. MiR-125a/b expressions and inflammatory cytokines were detected by RT-qPCR and ELISA assays in plasma samples from 120 sepsis patients. Besides, blood biochemical indexes, disease severity scores, and in-hospital mortality of sepsis patients were recorded. Meanwhile, miR-125a/b expressions in plasma from 120 health controls (HCs) were also detected by RT-qPCR. MiR-125b was elevated in sepsis patients compared with HCs, and ROC curve revealed that miR-125b could well distinguish sepsis patients from HCs with AUC 0.658. MiR-125b positively correlated with APACHE II score, SOFA score, Scr, CRP, PCT, TNF-α, and IL-6 levels. Most interestingly, miR-125b was greatly decreased in survivors compared with non-survivors, and multivariate analysis revealed that miR-125b independently predicted higher mortality risk in sepsis patients. Besides, miR-125a showed no significant correlation with sepsis risk, disease severity, or prognosis. MiR-125b but not miR-125a is upregulated and exhibits a trend to correlate with enhanced disease severity, inflammation, and increased mortality in sepsis patients.


[Cardiotropic effects of luliberin. Interaction with rat heart sarcolemmal membrane].
Studies of the effects of luliberin (luteinizing hormone-releasing hormone) on cardiac cells showed that the hormone is specifically bound by the sarcolemmal membrane. The activity of the membrane enzyme -- reception of luliberin involves two types of binding sites with high (K' = 10(7) M-1) and low (K'' = 10(5) M-1) affinities for the hormone, respectively. The hormone has no effect on the activities of adenylate cyclase and phosphodiesterase.


Simple synthesis of chitosan/alginate/graphene oxide/UiO-67 amphoteric aerogels: Characterization, adsorption mechanism and application for removal of cationic and anionic dyes from complex dye media.
A chitosan/alginate/graphene oxide/UiO-67 (CS/SA/GO/UiO-67) amphoteric aerogel was synthesized successfully. A series of characterization experiments of CS/SA/GO/UiO-67 amphoteric aerogel was performed by SEM, EDS, FT-IR, TGA, XRD, BET, and zeta potential. The competitive adsorption properties of different adsorbents for complex dyes wastewater (MB and CR) at room temperature (298 K) were compared. Langmuir isotherm model predicted that the maximum adsorption quantity of CS/SA/GO/UiO-67 for CR and MB was 1091.61 and 1313.95 mg/g, respectively. The optimum pH values of CS/SA/GO/UiO-67 for the adsorption of CR and MB were 5 and 10, respectively. The kinetic analysis showed that the adsorption of MB and CR on CS/SA/GO/UiO-67 was more suitable for the pseudo-second-order and pseudo-first-order kinetic model, respectively. The isotherm study revealed that the adsorption of MB and CR was consistent with the Langmuir isotherm model. The thermodynamic study demonstrated that the adsorption process of MB and CR was exothermic and spontaneous. FT-IR analysis and zeta potential characterization experiments revealed that the adsorption mechanism of MB and CR on CS/SA/GO/UiO-67 depended on π-π bond, hydrogen bond, and electrostatic attraction. Repeatable experiments showed that the removal rates of MB and CR of CS/SA/GO/UiO-67 after six cycles of adsorption were 67.19 and 60.82 %, respectively.


Lithium contamination of honeybee products and its accumulation in brood as a consequence of anti-varroa treatment.
Varroa destructor, the primary honeybee pathogen, is kept in check by various chemical compounds which may enter the human diet through honeybee products. Lithium is an emerging varroa control substance, and we investigated its accumulation in honey, bee bread, brood and adults along with the mortality of bees. Increased lithium concentrations were detected in workers, fed individually once per os with 10 µL of 25 mM LiCl in sucrose solution (6.50-40.10 mg/kg) or had the same solution available ad libitum (39.25-266.00 mg/kg). A three-day treatment of honeybee colonies with 25 mM LiCl in 1L/day sucrose solution increased lithium concentrations in five-day-old larvae, honey, and bee bread: up to 45.0, 1.2, and 47.0 mg/kg, respectively. Lithium concentrations peaked three days post-treatment in both larvae and honey and increased worker mortality was observed. The control colonies exhibited lithium concentrations below the limit of quantification (0.5 mg/kg). Prudence in lithium use is advised.


Faith-based partnerships in graduate medical education: the experience of the Morehouse School of Medicine Public Health/Preventive Medicine Residency Program.
Faith-based organizations can be strategic partners in addressing the needs of low-income and underserved individuals and communities. The Morehouse School of Medicine (MSM) Public Health/Preventive Medicine Residency Program (PH/PMR) collaborates with faith-based organizations for the purpose of resident education, community engagement, and service. These partners provide guidance for the program's community initiatives and health promotion activities designed to address health inequities. Residents complete a longitudinal community practicum experience with a faith-based organization over the 2-year training period. Residents conduct a community health needs assessment at the organization and design a health intervention that addresses the identified needs. The faith-based community practicum also serves as a vehicle for achieving skills in all eight domains of the Public Health Competencies developed by the Council on Linkages and all six Accreditation Council for Graduate Medical Education (ACGME) Core Competencies. The MSM PH/PMR Program has engaged in faith-based partnerships for 7 years. This article discusses the structure of these partnerships, how partners are identified, funding sources for supporting resident projects, and examples of resident health needs assessment and intervention activities. The MSM PH/PMR Program may serve as a model to other residency and fellowship programs that may have an interest in developing partnerships with faith-based organizations.


Application of capillary electrophoretic methods for the analysis of plant phloem and xylem saps composition: A review.
Plant vascular tissue is essential for the exchange of water, nutrients, metabolic products, and signals among distant organs in cormophytes. The compositions of phloem and xylem saps are highly dependent on many internal and external factors, and thus their analysis provides a valuable insight into plant physiology, growth, and development as well as nutrition status or presence of biotic or abiotic stresses. Capillary electrophoresis characterized by highly efficient separations and minuscule sample requirements represents a suitable analytical technique for this purpose because the sap constitutes a complex mixture with generally minimal availability. This review aims at providing a comprehensive overview of published capillary electrophoretic methods for the analysis of primary components present in the phloem and xylem saps of higher plants.


Pulsatile release of biomolecules from polydimethylsiloxane (PDMS) chips with hydrolytically degradable seals.
We demonstrate, for the first time, a robust novel polydimethylsiloxane (PDMS) chip that can provide controlled pulsatile release of DNA based molecules, proteins and oligonucleotides without external stimuli or triggers. The PDMS chip with arrays of wells was constructed by replica molding. Poly(lactic acid-co-glycolic acid) (PLGA) polymer films of varying composition and thickness were used as seals to the wells. The composition, molecular weight and thickness of the PLGA films were all parameters used to control the degradation rate of the seals and therefore the release profiles. Degradation of the films followed the PLGA composition order of 50:50 PLGA>75:25 PLGA>85:15 PLGA at all time-points beyond week 1. Scanning electron microscopy images showed that films were initially smooth, became porous and ruptured as the osmotic pressure pushed the degrading PLGA film outwards. Pulsatile release of DNA was controlled by the composition and thickness of the PLGA used to seal the well. Transfection experiments in a model Human Embryonic Kidney 293 (HEK293) cell line showed that plasmid DNA loaded in the wells was functional after pulsatile release in comparison to control plasmid DNA at all time-points. Thicker films degraded faster than thinner films and could be used to fine-tune the release of DNA over day length periods. Finally the PDMS chip was shown to provide repeated sequential release of CpG oligonucleotides and a model antigen, Ovalbumin (OVA), indicating significant potential for this device for vaccinations or applications that require defined complex release patterns of a variety of chemicals, drugs and biomolecules.


Vertical fluctuations of phospholipid acyl chains in bilayers.
The possibility of vertical displacement of acyl chains in lipid bilayers has been examined by quenching the fluorescence of 2-(9-anthroyloxy)palmitic acid with 5- and 16-doxylstearates in dipalmitoylphosphatidylcholine unilamellar vesicles. Measurement of lifetime and steady-state quenching showed that the dynamic component of quenching was independent of the transverse position of the quencher indicating that a quencher at the 16-position could interact with a fluorophore at the 2-position with high frequency. The differences in steady-state quenching could be accounted for by the differences in the static component of quenching. The results provide further evidence for rapid vertical displacements of acyl chains in phospholipid bilayers.


Identification of regions within the Legionella pneumophila VipA effector protein involved in actin binding and polymerization and in interference with eukaryotic organelle trafficking.
The Legionella pneumophila effector protein VipA is an actin nucleator that co-localizes with actin filaments and early endosomes in infected macrophages and which interferes with organelle trafficking when expressed in yeast. To identify the regions of VipA involved in its subcellular localization and functions, we ectopically expressed specific VipA mutant proteins in eukaryotic cells. This indicated that the characteristic punctate distribution of VipA depends on its NH2 -terminal (amino acid residues 1-133) and central coiled-coil (amino acid residues 133-206) regions, and suggested a role for the COOH-terminal (amino acid residues 206-339) region in association with actin filaments and for the NH2 -terminal in co-localization with early endosomes. Co-immunoprecipitation and in vitro assays showed that the COOH-terminal region of VipA is necessary and sufficient to mediate actin binding, and is essential but insufficient to induce microfilament formation. Assays in yeast revealed that the NH2 and the COOH-terminal regions, and possibly an NPY motif within the NH2 region of VipA, are necessary for interference with organelle trafficking. Overall, this suggests that subversion of eukaryotic vesicular trafficking by VipA involves both its ability to associate with early endosomes via its NH2 -terminal region and its capacity to bind and polymerize actin through its COOH-terminal region.


Characterization and functional properties of mango peel pectin extracted by ultrasound assisted citric acid.
Pectin was extracted from 'Tainong No. 1' mango peels, using a chelating agent-citric acid as extraction medium by ultrasound-assisted extraction (UAE) and conventional extraction (CE) at temperatures of 20 and 80°C. Chemical structures, rheological and emulsifying properties of mango peel pectins (MPPs) were comparatively studied with laboratory grade citrus pectin (CP). All MPPs exhibited higher protein content (4.74%-5.94%), degree of methoxylation (85.43-88.38%), average molecular weight (Mw, 378.4-2858kDa) than the CP, but lower galacuronic acid content (GalA, 52.21-53.35%). CE or UAE at 80°C resulted in significantly higher pectin yield than those at 20°C, while the extraction time for UAE-80°C (15min) was significantly shorter compared to CE-80°C (2h) with comparable pectin yield. Moreover, MPPs extracted at 80°C were observed with higher GalA and protein content, higher Mw, resulting in higher viscosity, better emulsifying capacity and stability, as compared to those extracted at 20°C and the CP. Therefore, these results suggested that MPPs from 'Tainong No. 1' may become a highly promising pectin with good thickening and emulsifying properties, using ultrasound-assisted citric acid as an efficient and eco-friendly extraction method.


Combinations of multivariate statistical analysis and analytical hierarchical process for indexing surface water quality under arid conditions.
Novel methods for water quality indexing increase insight into the fitness of water bodies for different uses. We hypothesized that integrating multivariate statistical analysis (MSA) with the analytical hierarchical process (AHP) may provide a reliable estimation of water quality status. Hence, twenty water samples from canals and drains in the northern Nile Delta, Egypt were collected during summer, autumn, winter, and spring and analyzed. Data were subjected to MSA, including correlation analysis, principal component analysis (PCA), and hierarchal cluster analysis (HCA). The AHP was applied to derive weights of parameters implied in developing water quality indices for irrigation (IWQI) and fish farming (FFWQI). Human health risks due to exposure to potentially toxic elements (PTEs) via dermal contact were also considered. The average concentrations of water constituents were acceptable for irrigation, except sodium adsorption ratio (SAR) and Cl-. The dissolved oxygen, total dissolved solids, Cl-, NO2-N, NO3-N, NH3, and PTEs (except Zn) did not meet standard limits for fish production. The MSA revealed that water contamination resulted from human activities (agriculture, industry, and domestic wastes) and hydrochemical processes. The PCA indicated that SAR, Cu, and pH could adequately represent water quality for irrigation, while temperature, NO2-N, Cr, and Zn could reflect fish farming requirements. The AHP provided consistent weights for the original and shortlisted parameters. The water quality varied from good to poor for irrigation and from excellent to low for fish farming. The minimum IWQI could adequately represent the IWQI (R2 = 0.83) and thus reduce the time, effort, and cost for monitoring water quality. However, the minimum FFWQI showed moderate consistency (R2 = 0.51) with FFWQI, implying that increasing the sampling size is essential for better performance. The hazard quotient of all PTEs was below 1.0 for both adults and children, indicating a safe limit. The potential cancer risk was acceptable (1.36E-06) for adults and safe (8.03E-07) for children. Results of this work would be a start point for efficient quality control programs in arid regions.


Circulating Monocytes in between the Gut and the Mind.
Hippocampal neurogenesis is affected throughout life by factors external to the brain, including what we eat, our gut microbiota, and the immune system. However, the mechanisms that link microbiota to neurogenesis are still puzzling. Now in Cell Reports, Möhle et al. (2016) attribute a role to Ly6C(hi) monocytes in this gut-immune-brain axis.


Repeated donkey milk consumption reduces anxiety-like behaviors and brain oxidative damage to lipids in mice.
Donkey milk (DM) is a source of bioactive compounds that can benefit neural functioning. In the present study, we investigated the effects of DM consumption on anxiolytic-related, despair-like, locomotion and coordination behaviors, as well as the provision of protection from oxidative damage to lipids and proteins in brain tissues and melatonin plasma levels. To achieve this, male mice orally received DM (4 g.kg-1) or vehicle for 18 days. Their behavior was assessed in the following tests: elevated plus maze (EPM), open field and rotarod tests (OF, RR) and forced swimming test (FST). Acute treatments with diazepam (DZP, 1.5 mg.kg-1, v.o.), fluoxetine (FLX, 20 mg.kg-1, i.p.) and nortriptyline (NTP, 20 mg.kg-1, i.p.) were used as positive controls. On the eighteenth day, the animals were euthanized and brain tissue and blood were collected to measure oxidative damage, and melatonin plasma levels. Similar to DZP, repeated DM consumption reduced exploration to open areas in the EPM test. Under our experimental conditions, conventional antidepressants reduced immobility time in the FST, and the benzodiazepine treatment impaired motor coordination in mice. No significant differences in locomotion, motor coordination and despair-related behaviors were observed in the mice treated with DM when assessed in the EPM, OF, RR and FST, respectively. Biochemical assays showed that repeated DM exposition protected against oxidative damage to lipids and increased plasma levels of melatonin. These findings suggest consumption of DM may be a promising food for the treatment of anxiety-related disorders, without depressant effects on the central nervous system.


A deletion mutation in Slc12a6 is associated with neuromuscular disease in gaxp mice.
Giant axonopathy (gaxp), an autosomal recessive mouse mutation, exhibits ataxia of the hind legs with a slight side-to-side wobble while walking. Within the genomic region of the gaxp locus, a total of 94 transcripts were identified; the annotation of these genes using OMIM and PubMed yielded three potential candidate genes. By cDNA microarray analysis, 54 genes located on or near the gaxp locus were found to exhibit differential expression between gaxp and littermate controls. Based on microarray data and the known function of genes identified, Slc12a6 was selected as the primary candidate gene and analyzed using the Reveal technology of SpectruMedix. A 17-base deletion was detected from within exon 4 of Slc12a6. Reverse transcriptase polymerase chain reaction validated the difference in Slc12a6 expression in different types of mice at the mRNA level, revealing a marked reduction in gaxp mice. Western blot analysis indicated that the protein product of Slc12a6, the K(+)-Cl(-) cotransporter Kcc3, was not detectable in gaxp mice. The causative role of the exon 4 mutation within Slc12a6 in the gaxp phenotype was further confirmed by screening multiple inbred strains and by excluding the mutation of nearby genes within the gaxp locus.


Microbial transformation of nandrolone decanoate by acremonium strictum.
Estr-4-en-3,17-dione II, 17beta-hydroxyestr-4-en-3-one III, 15alpha-hydroxyestr-4-en-3,17-dione IV, and 15alpha,17beta-dihydroxyestr-4-en-3-one V were produced by microbial transformation of nandrolone decanoate I in the culture of Acremonium strictum PTCC 5282. Bioconversion characteristics observed were ester hydrolysis, oxidation, and hydroxylation. Each microbial product was purified chromatographically and characterized on the basis of spectral data obtained from (1)H-NMR,( 13)C-NMR, FT-IR, MS, and physical constants such as melting point and optical rotation.


Refractory headache in a patient with breast cancer and carcinomatous meningitis unresponsive to analgesics: case report.
A 44-year old female presented with locally advanced breast cancer that had been treated with neoadjuvant chemotherapy followed by modified radical mastectomy and thereafter three cycles of paclitaxel. She presented with severe refractory headache that was unresponsive to oral analgesics including morphine. Both CT and MRI scans with contrast were normal, however her cerebrospinal fluid was positive for malignant cells. The patient's headache responded to whole brain radiotherapy.


Hormone-induced subcellular redistribution of trimeric G proteins.
Trimeric guanine nucleotide-binding proteins (G proteins) function as the key regulatory elements in a number of transmembrane signaling cascades where they convey information from agonist-activated receptors to effector molecules. The subcellular localization of G proteins is directly related to their functional role, i.e., the dominant portion of the cellular pool of G proteins resides in the plasma membrane. An intimate association of G protein subunits with the plasma membrane has been well known for a long time. However, results of a number of independent studies published in the past decade have indicated clearly that exposure of intact target cells to agonists results in subcellular redistribution of the cognate G proteins from plasma membranes to the light-vesicular membrane fractions, in internalization from the cell surface into the cell interior and in transfer from the membrane to the soluble cell fraction (high-speed supernatant), i.e., solubilization. Solubilization of G protein a subunits as a consequence of stimulation of G protein-coupled receptors (GPCRs) with agonists has also been observed in isolated membrane preparations. The membrane-cytosol shift of G proteins was detected even after direct activation of these proteins by non-hydrolyzable analogues of GTP or by cholera toxin-induced ADP-ribosylation. In addition, prolonged stimulation of GPCRs with agonists has been shown to lead to down-regulation of the relevant G proteins. Together, these data suggest that G proteins might potentially participate in a highly complex set of events, which are generally termed desensitization of the hormone response. Internalization, subcellular redistribution, solubilization, and down-regulation of trimeric G proteins may thus provide an additional means (i.e., beside receptor-based mechanisms) to dampen the hormone or neurotransmitter response after sustained (long-term) exposure.


Implant size and mechanical properties influence the failure of the adhesive bond between cartilage implants and native tissue in a finite element analysis.
Implanted cartilage replacements (ICRs) are a promising approach to restore the functionality of joints with partial- and full-thickness articular cartilage lesions. Two major hurdles hindering successful repair of cartilage injuries with ICRs are their inadequate mechanical properties and fixation into the defect area. While the ICR geometry and mechanical properties are expected to affect the loads and deformations at the adhesive interface between the implant and native cartilage, little is known about the relationship between these implant characteristics and the quality of the fixation. The objective of this study was to evaluate the effect of implant size, thickness, modulus, surface coefficient of friction and Poisson's ratio on the failure of a fibrin adhesive interface. These factors were evaluated in an idealized finite element model of the medial compartment of the human knee with the damage and failure of fibrin at the implant/cartilage interface represented by a cohesive zone model. Both axial compression and sliding were included in the loading conditions. The results demonstrated that ICR size and material properties have a significant effect on the failure of the fibrin that adheres the implant to the native tissue. Lack of anchorage to underlying bone, larger implant sizes, higher surface coefficient of friction and higher compliance of the implant can increase the chance of implant loosening and delamination. In the future, these results may guide implant design and cartilage repair techniques.


Circulating tumour DNA biomarkers in savolitinib-treated patients with non-small cell lung cancer harbouring MET exon 14 skipping alterations: a post hoc analysis of a pivotal phase 2 study.
Savolitinib, a selective MET inhibitor, showed efficacy in patients with non-small cell lung cancer (NSCLC), including pulmonary sarcomatoid carcinoma (PSC), harbouring MET exon 14 skipping alteration (METex14). To analyse post hoc, the association between circulating tumour DNA (ctDNA) biomarkers and clinical outcomes, including resistance, with savolitinib. A multicentre, single-arm, open-label phase 2 study. All enrolled patients with baseline plasma samples were included. Outcomes were objective response rate (ORR), progression-free survival (PFS) and overall survival (OS) by baseline METex14 and post-treatment clearance, coexisting gene alterations at baseline and disease progression. Among 66 patients with baseline ctDNA sequencing, 46 (70%) had detectable METex14. Frequent coexisting baseline gene alterations included TP53 and POT1 mutations. Patients with detectable baseline METex14 exhibited worse PFS [hazard ratio (HR), 1.77; 95% confidence interval (CI), 0.88-3.57; p = 0.108] and OS (HR, 3.26; 95% CI, 1.35-7.89; p = 0.006) than those without, despite showing a numerically higher ORR. Among 24 patients with baseline detectable METex14 and evaluable postbaseline samples, 13 achieved METex14 clearance post-treatment. Median time to first clearance was 1.3 months (range, 0.7-1.5). METex14 post-treatment clearance was associated with better ORR (92.3%; 95% CI, 64.0-99.8 versus 36.4%; 95% CI, 10.9-69.2; p = 0.0078), PFS (HR, 0.44; 95% CI, 0.2-1.3; p = 0.1225) and OS (HR, 0.31; 95% CI, 0.1-1.0; p = 0.0397) versus non-clearance. Among 22 patients with disease progression, 10 acquired pathway alterations (e.g. in RAS/RAF and PI3K/PTEN) alone or with secondary MET mutations (D1228H/N and Y1230C/H/S). ctDNA biomarkers may allow for longitudinal monitoring of clinical outcomes with savolitinib in patients with METex14-positive PSC and other NSCLC subtypes. Specifically, undetectable baseline METex14 or post-treatment clearance may predict favourable clinical outcomes, while secondary MET mutations and other acquired gene alterations may explain resistance to savolitinib. The trial was registered with ClinicalTrials.gov (NCT02897479) on 13 September 2016.


Cross-sectional imaging of thoracic and abdominal complications of cerebrospinal fluid shunt catheters.
This study aims to review the imaging findings of distal (thoracic and abdominal) complications related to ventriculo-peritoneal (VP), ventriculo-pleural (VPL), and ventriculo-atrial (VA) cerebrospinal fluid (CSF) shunt catheter placement. Institution review board-approved single-center study of patients with thoracic and abdominal CSF catheter-related complications on cross-sectional imaging examinations over a 14-year period was performed. Clinical presentation, patient demographics, prior medical history, and subsequent surgical treatment were recorded. The presence or absence of CSF catheter-related infection and/or acute hydrocephalus on cross-sectional imaging was also recorded. There were 81 distal CSF catheter-related complications identified on 47 thoracic or abdominal imaging examinations in 30 patients (age 5-80 years, mean 39.3 years), most often on CT (CT = 42, MRI = 1, US = 4). Complications included 38 intraperitoneal and 11 extraperitoneal fluid collections. Extraperitoneal collections included nine abdominal wall subcutaneous (SC) pseudocysts associated with shunt migration and obesity, an intrapleural pseudocyst, and a breast pseudocyst. There were also two large VPL-related pleural effusions, a fractured catheter in the SC tissues, and a large VA shunt thrombus within the right atrium. Ten patients (33.3 %) had culture-positive infection from CSF or shunt catheter samples. Ten patients (33.3 %) had features of temporally related acute or worsening hydrocephalus on neuroimaging. In four of these patients, the detection of thoracic and abdominal complications on CT preceded and predicted the findings of acute hydrocephalus on cranial imaging. Thoracic and abdominal complications of CSF shunts, as can be identified on CT, include shunt infection and/or obstruction, may be both multiple and recurrent, and may be predictive of concurrent acute intracranial problems.


Beta interferons from the extant camelids: Unique among eutherian mammals.
The COVID-19 pandemic is a wake-up call on the zoonotic viral spillover events and the need to be prepared for future outbreaks. Zoonotic RNA viruses like the Middle East respiratory syndrome coronavirus (MERS-CoV) are potential pathogens that could trigger the next pandemic. Dromedary camels are the only known animal source of MERS-CoV zoonotic infections, but little is known about the molecular antiviral response in this species. IFN-β and other type-I interferons provide the first line of defense against invading pathogens in the host immune response. We identified the IFNB gene of the dromedary camel and all extant members of the family Camelidae. Camelid IFN-β is unique with an even number of cysteines in the mature protein compared to other eutherian mammals with an odd number of cysteines. The viral mimetic poly(I:C) strongly induced IFN-β expression in camel kidney cells. Induction of IFN-β expression upon infection with camelpox virus was late and subdued when compared to poly(I:C) treatment. Prokaryotically expressed recombinant dromedary IFN-β induced expression of IFN-responsive genes in camel kidney cells. Further, recombinant IFN-β conferred antiviral resistance to camel kidney cells against the cytopathic effects of the camelpox virus, an endemic zoonotic pathogen. IFN-β from this unique group of mammals will offer insights into antiviral immune mechanisms and aid in the development of specific antivirals against pathogens that have the potential to be the next zoonotic pandemic.


Field analyses of insect heat budgets: Reflectance, size and heating rates.
This paper outlines simple techniques for determining rates of heat gain and loss in relation to the weight and reflectance of insects caught in their natural habitats. In particular the construction of a new 'reflectometer' is described. The results thus obtained permit estimates of the relative importance of size and colour in determining rates of heat exchange and temperature excesses, so allowing better predictions of heat budgets for a given species.


Acute kidney injury risk in orthopaedic trauma patients pre and post surgery using a biomarker algorithm and clinical risk score.
Acute kidney injury (AKI) after major trauma is associated with increased mortality. The aim of this study was to assess if measurement of blood biomarkers in combination with clinical characteristics could be used to develop a tool to assist clinicians in identifying which orthopaedic trauma patients are at risk of AKI. This is a prospective study of 237 orthopaedic trauma patients who were consecutively scheduled for open reduction and internal fixation of their fracture between May 2012 and August 2013. Clinical characteristics were recorded, and 28 biomarkers were analysed in patient blood samples. Post operatively a combination of H-FABP, sTNFR1 and MK had the highest predictive ability to identify patients at risk of developing AKI (AUROC 0.885). Three clinical characteristics; age, dementia and hypertension were identified in the orthopaedic trauma patients as potential risks for the development of AKI. Combining biomarker data with clinical characteristics allowed us to develop a proactive AKI clinical tool, which grouped patients into four risk categories that were associated with a clinical management regime that impacted patient care, management, length of hospital stay, and efficient use of hospital resources.