Long-Acting Efavirenz and HIV-1 Fusion Inhibitor Peptide Co-loaded Polymer-Lipid Hybrid Nanoparticles: Statistical Optimization, Cellular Uptake, and In Vivo Biodistribution.
The objective of the present study was to develop long-acting efavirenz (Efa)-enfuvirtide (Enf) Co-loaded polymer-lipid hybrid nanoparticles (PLN) with improved intracellular delivery to target T-cells and macrophage cells located in multiple human immunodeficiency virus sanctuaries. The Box-Behnken design was utilized to optimize three high-risk factors, namely, Efa amount, sonication time for primary emulsion, and sonication time for aqueous nanodispersion obtained from preliminary studies. Lyophilized Efa-Enf Co-loaded PLN using trehalose elicited spherical morphology, drug amorphization on incorporation, and absence of drug-excipient interaction. In vitro release studies revealed an sustained release of both the drugs from PLN with the differential release profile. Efa-Enf Co-loaded PLN exhibited low hemolytic, platelet and leukocyte aggregation as well as low cytotoxicity in Jurkat E6.1 T-cells and U937 macrophage cells. Circular dichroism spectra confirmed the presence of an α-helix form of Enf after encapsulation in PLN. Coumarin-6-loaded PLN exhibited enhanced cellular uptake in Jurkat E6.1 T-cells and U937 macrophage cells in comparison to free coumarin-6, as evidenced by fluorescence microscopy and flow cytometry. In vivo biodistribution studies after intravenous administration of near-infrared dye-loaded PLN (surrogate for Efa-Enf PLN) revealed non-uniform distribution within 2 h in the order of spleen ≥ liver > lymph node > thymus > lungs > female reproductive tract (FRT) > heart > kidneys > brain. However, subcutaneous administration caused non-uniform biodistribution after 3 days, eliciting a long-acting slow release from the injection site depot until day 5 in the infection-spread site (lymph nodes and FRT), reservoir sites (liver and spleen) and the difficult-to-access site (brain). Furthermore, it presents a vital illustration of the available tissue-specific drug concentration prediction from simulated surrogate PLN.


Genetic studies on the Gc subtypes.
Gc subtypes were determined by immunofixation isoelectric focusing. In a family study involving 93 families with a total of 176 children, a three-allele model with six genotypes was confirmed. The distribution of Gc subtypes was studied in three populations from Europe and in a sample of Druzes from Northern Israel. Three new variant Gc types were observed.


Plasmodium falciparum antioxidant protein as a model enzyme for a special class of glutaredoxin/glutathione-dependent peroxiredoxins.
Peroxiredoxins are important heterogeneous thiol-dependent hydroperoxidases with a variety of isoforms and enzymatic mechanisms. A special subclass of glutaredoxin/glutathione-dependent peroxiredoxins has been discovered in bacteria and eukaryotes during the last decade, but the exact enzymatic mechanisms of these enzymes remain to be unraveled. We performed a comprehensive analysis of the enzyme kinetics and redox states of one of these glutaredoxin/glutathione-dependent peroxiredoxins, the antioxidant protein from the malaria parasite Plasmodium falciparum, using steady-state kinetic measurements, site-directed mutagenesis, redox mobility shift assays, gel filtration, and mass spectrometry. P. falciparum antioxidant protein requires not only glutaredoxin but also glutathione as a true substrate for the reduction of hydroperoxides. One peroxiredoxin cysteine residue and one glutaredoxin cysteine residue are sufficient for catalysis, however, additional cysteine residues of both proteins result in alternative redox states and conformations in vitro with implications for redox regulation. Our data furthermore point to a glutathione-dependent peroxiredoxin activation and a negative subunit cooperativity. The investigated glutaredoxin/glutathione/peroxiredoxin system provides numerous new insights into the mechanism and redox regulation of peroxiredoxins. As a member of the special subclass of glutaredoxin/glutathione-dependent peroxiredoxins, the P. falciparum antioxidant protein could become a reference protein for peroxiredoxin catalysis and regulation.


The effect of aerobic exercise on tumour blood delivery: a systematic review and meta-analysis.
Tumour blood vessels are structurally and functionally abnormal, resulting in areas of hypoxia and heterogeneous blood supply. Aerobic exercise may modulate tumour blood flow and normalise the tumour microenvironment to improve chemotherapy delivery. This systematic review and meta-analysis aimed to evaluate the effect of the aerobic exercise mode on tumour hypoxia, vascularisation and blood flow. Four online databases were searched. Preclinical and clinical randomised controlled trials examining the effects of aerobic exercise training on hypoxia, vascularisation or blood flow in solid tumours were included. The risk of bias was assessed and a meta-analysis performed. Seventeen preclinical studies and one clinical study met criteria. Eleven studies assessed hypoxia, 15 studies assessed vascularisation and seven evaluated blood flow. There was large variability in measurement methods, tumour types and exercise program designs. The overall risk of bias was unclear in clinical and preclinical studies, owing to poor reporting. There was no significant effect of aerobic exercise on hypoxia (SMD = -0.17; 95% CI = -0.62, 0.28; I2 = 60%), vascularisation (SMD = 0.07; 95% CI = -0.40, 0.55; I2 = 71%) or blood flow (SMD = 0.01; 95% CI = -0.59, 0.61; I2 = 63%). There is heterogeneity in methodology, resulting in evidence that is inconsistent and inconclusive for the effects of aerobic exercise on hypoxia, vascularisation and blood flow. Most evidence of aerobic exercise effects on tumour blood flow is in animal models, with very limited evidence in humans.


Effect of arginine vasopressin on coronary and systemic hemodynamics in man.
Arginine vasopressin is a potent constrictor of isolated arterial segments in vitro. However, it is disputed whether, in vivo, arginine vasopressin acts as either a systemic or coronary vasoconstrictor. The purpose of this study was to determine the effect of arginine vasopressin on coronary and systemic vascular tone in man. Six patients undergoing routine diagnostic cardiac catheterization were studied. At angiography, all patients were found to have severe coronary artery disease. Cardiac and systemic pressures and coronary blood flow as well as serum arginine vasopressin and osmolality levels were measured before and after administration of hypertonic contrast dye and then following intravenous injection of an analog inhibitor of arginine vasopressin. At baseline, serum arginine vasopressin levels were within the normal range in all patients, but, following infusion of contrast dye, rose above physiological levels in 5 patients. In these latter patients, there were no significant changes in coronary and systemic hemodynamics between baseline (condition 1), following administration of contrast dye (condition 2) and 10 minutes after injection of arginine vasopressin inhibitor (condition 3). Mean coronary sinus blood flow (ml/min) and mean coronary vascular resistance (dyne.sec.cm-5) were 184 +/- 49 and 41,235 +/- 8275 during condition 1, 204 +/- 62 and 39,442 +/- 8781 during condition 2, and 192 +/- 75 and 44,930 +/- 11455 during condition 3. Mean arterial pressure (mm Hg) and mean systemic vascular resistance (dyne.sec.cm-5) during conditions 1, 2 and 3 were 105 +/- 12 and 1454 +/- 174, 114 +/- 21 and 1432 +/- 210, and 114 +/- 15 and 1436 +/- 147, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Terrestrial humic-like fluorescence peak of chromophoric dissolved organic matter as a new potential indicator tracing the antibiotics in typical polluted watershed.
Natural surface waters are threatened globally by antibiotics pollution. In this study, we analyzed antibiotics and CDOM (Chromophoric dissolved organic matter) fluorescence in different water bodies using HPLC method and Excitation Emission Matrix- Parallel factor analysis, respectively. A combination of field studies in the Yinma River Watershed were conducted in rivers, reservoirs and urban rivers, and 65 CDOM and antibiotic samples were taken in April, May, July, and August 2016. EEM-PARAFAC analysis identified two components; a humic-like (C1) component and a tryptophan-like (C2) component. The redundancy analysis (RDA) demonstrated that CDOM could explain 38.2% (two axes) of the five antibiotics in reservoirs (N = 31), and 26.0% (two axes) of those in rivers and urban water (N = 30). Furthermore, the Pearson correlation coefficient between Sulfamethoxazole and C1 in reservoir water was 0.91 (t-test, 2-tailed, p < 0.01), and that between Sulfamethoxazole and C2 was 0.68 (t-test, 2-tailed, p < 0.01). This indicated that the humic-like component of CDOM PARAFAC fluorescence could detect Sulfamethoxazole contamination levels in the homogenized reservoir waters. Our results identified Sulfamethoxazole and Quinolones (Norfloxacin, 16.5 ng L-1; Enrofloxacin, 0.3 ng L-1; Ciprofloxacin, 30.9 ng L-1) at mean concentrations of 369.5 ng L-1 and 15.9 ng L-1, respectively, which were the higher levels in natural surface waters. The FTIR spectroscopy of the mixture of humic acid and sulfamethoxazole showed that the absorbance at 3415 cm-1 linked to OH stretching of OH groups and at 1386 cm-1 because of OH bending and vibration of COOH groups became weaker, indicating that COOH groups of humic acid can adsorb and react with -NH2 of sulfamethoxazole. The CDOM PARAFAC components can be adapted for online or in situ fluorescence measurements as an early warning of Sulfamethoxazole distribution and contamination in similar aquatic environments.


Susceptibility and intra-species transmission of the H9N2 G1 prototype lineage virus in Japanese quail and turkeys.
Avian influenza viruses of the H9N2 subtype have circulated in the poultry population in Asia, Far and Middle East since the mid-1990 s. One of the most widespread lineages established in poultry is the G1 lineage. This lineage has undergone further evolution and reassortment since its first detection in 1997 and G1-like H9N2 viruses still circulate. In this study we have investigated the susceptibility of quail and turkeys to the H9N2 G1-lineage prototype strain (A/quail/Hong Kong/G1/97). Contact transmission experiments were carried out in both avian species. Animals were infected oro-nasally with increasing doses of the virus (10(3)-10(6) EID 50/0.1 ml) and sentinel birds were introduced 4 days post infection (pi) in each experimental group. Quail were more susceptible than turkeys, as they were readily infected with lower challenge doses. Interestingly, infection of turkeys was associated with worse clinical condition. Transmission was detected in both species. Quail infected with a dose less than or equal to 10(4) EID50 transmitted the virus to the sentinels without showing any signs of disease. These findings reinforce the hypothesis that quail may ensure the perpetuation of H9N2 viruses in poultry, acting as a silent reservoir.


The perception threshold to an electric stimulus deeply applied in the lower limbs in normal and diabetic subjects.
The loss of sensitivity to nociceptive stimuli is one of the main factors involved in the pathogeny of diabetic gangrene. The aim of this study was to develop a simple and practical method for selecting the cases prone to such a complication. The perception threshold (either voltage, mV or current, microA) was determined in 137 randomly selected diabetic patients and 38 non-diabetic controls, by an original electronic device using sinusoidal waves, delivered through two electrode needles introduced at a depth of 0.5 cm at two points located on the anterior aspect of the shank about 12 cm apart. At each of the eight frequencies studied (10, 20, 50, 100, 200, 500, 1000 and 2000 Hz) three electrical parameters (voltage across the needle electrodes, the resistive and the capacitive current component) were recorded when the subject first perceived the stimulus. A significantly higher perception threshold was found in diabetic patients vs. non-diabetic subjects, at all frequencies studied and with all three electrical parameters recorded. A greater difference in the perception threshold between diabetic and non-diabetic subjects (a ratio higher than 3) was, however, found using the resistive component of the current and at frequencies higher than 200 Hz.


A service system planning model for individuals with serious mental illness.
An institutional-based care system in mental health has been replaced by a network of community-based services with different levels of structure and support. This poses both an opportunity and a challenge to provide appropriate and effective care to persons with serious mental illnesses. This paper describes a simulation-based approach for mental health system planning, focused on hospital and residential service components that can be used as a decision support tool. A key feature of this approach is the ability to represent the current service configuration of psychiatric care and the client flow pattern within that framework. The strength of the simulation model is to help mental health service managers and planners visualize the interconnected nature of client flow in their mental health system and understand possible impacts of changes in arrival rates, service times, and bed capacity on overall system performance. The planning model will assist state mental health agencies to respond to requirements of the Olmstead decision to ensure that individuals with serious mental illness receive care in the least restrictive setting. Future plans for refining the model and its application to other service systems is discussed.


Incidence and Risk Factors for Nontuberculous Mycobacterial Infection after Allogeneic Hematopoietic Cell Transplantation.
Allogenic hematopoietic stem cell transplant (HCT) recipients are at risk of many infections. Nontuberculous mycobacteria (NTM) are increasingly recognized as clinically significant pathogens in this population. We investigated the incidence and risk factors for NTM infection after allogeneic HCT. This retrospective cohort study included all patients with allogeneic HCT at our institution during 2001 to 2013. Patients who developed significant NTM infection (NTM disease) were identified. Multivariable modeling was used to identify risk factors for NTM disease, and a risk score model was constructed to identify high-risk patients. Of 1097 allogeneic HCT patients, 45 (4.1%) had NTM isolated and 30 (2.7%) had NTM disease (28 [93.3%] exclusively pulmonary, 2 [6.7%] pulmonary plus another site). Incidence of NTM infection by competing risk analysis was 2.8% at 5 years (95% CI, 1.9% to 4.0%). The median time to diagnosis was 343 days (range, 19 to 1967). In Fine-Gray proportional hazards modeling, only global severity of chronic graft-versus-host disease (cGVHD) (HR, 1.99; 95% CI, 1.12 to 3.53; P = .019,) and cytomegalovirus (CMV) viremia (HR, 5.77; 95% CI, 1.71 to 19.45; P = .004) were significantly associated with NTM disease. Using these variables a risk score was calculated: 1 point for CMV viremia or moderate cGVHD and 2 points for severe cGVHD. The score divided patients into low risk (0 to 1 points, n = 820 [77.3%], 3-year NTM risk 1.2%), intermediate risk (2 points, n = 161 [15.4%], 3-year NTM risk 7.1%), and high risk (3 points, n = 56 [5.4%], 3-year NTM risk 14.3%). NTM disease after allogeneic HCT is common. Severe cGVHD and CMV viremia are associated with increased risk, permitting risk stratification.


Metabolic engineering of Escherichia coli for improved 6-deoxyerythronolide B production.
Escherichia coli is an attractive candidate as a host for polyketide production and has been engineered to produce the erythromycin precursor polyketide 6-deoxyerythronolide B (6dEB). In order to identify and optimize parameters that affect polyketide production in engineered E. coli, we first investigated the supply of the extender unit ( 2S)-methylmalonyl-CoA via three independent pathways. Expression of the Streptomyces coelicolor malonyl/methylmalonyl-CoA ligase ( matB) pathway in E. coli together with methylmalonate feeding resulted in the accumulation of intracellular methylmalonyl-CoA to as much as 90% of the acyl-CoA pool. Surprisingly, the methylmalonyl-CoA generated from the matB pathway was not converted into 6dEB. In strains expressing either the S. coelicolor propionyl-CoA carboxylase (PCC) pathway or the Propionibacteria shermanii methylmalonyl-CoA mutase/epimerase pathway, methylmalonyl-CoA accumulated up to 30% of the total acyl-CoA pools, and 6dEB was produced; titers were fivefold higher when strains contained the PCC pathway rather than the mutase pathway. When the PCC and mutase pathways were expressed simultaneously, the PCC pathway predominated, as indicated by greater flux of (13)C-propionate into 6dEB through the PCC pathway. To further optimize the E. coli production strain, we improved 6dEB titers by integrating the PCC and mutase pathways into the E. coli chromosome and by expressing the 6-deoxyerythronolide B synthase (DEBS) genes from a stable plasmid system.


Prevalence and characteristics of attaching and effacing strains of Escherichia coli isolated from diarrheic and healthy sheep and goats.
To determine the prevalence and characteristics of attaching and effacing Escherichia coli (AEEC) in diarrheic and healthy small ruminants. 502 lambs and kids with diarrhea and 511 healthy sheep and goats. Fecal samples from diarrheic and healthy sheep and goats were screened for the eae gene. In addition, E coli isolates with positive results for the eae gene (E coli eae+) were analyzed for the espB gene, production of verotoxins (VT), and serogroup. A significantly higher prevalence of healthy lambs and kids were infected with AEEC, compared with diarrheic lambs and kids and healthy adult sheep and goats. Some differences in the characteristics of E coli eae strains isolated from diarrheic and healthy animals were detected. Thus, the espB gene was detected more frequently among E coli eae+ strains isolated from healthy animals than in those isolated from diarrheic animals, and VT production was only detected in E coli eae+ strains isolated from healthy lambs and kids. The E coli eae+ isolates belonged to several O serogroups. However, 17 of 40 (42.5%) isolates from diarrheic lambs and only 4 of 168 (2.4%) isolates from healthy sheep belonged to serogroup 026. Our results suggest that E coli eae+ 026 strains may play a role in diarrheal disease in lambs, whereas E coli eae+ strains that also had VT production and eae+ strains that had positive results for the espB gene did not appear to be associated with diarrhea in small ruminants.


Outcomes of incontinent ileovesicostomy in the pediatric patient.
Ileovesicostomy is a reconstructive option in complex urological cases but pediatric specific outcomes are lacking. We report our results with pediatric ileovesicostomy. We retrospectively evaluated patients younger than 18 years undergoing incontinent ileovesicostomy at Vanderbilt University. History, urinary tract management and operative course were reviewed in the electronic medical record. Particular attention was given to immediate and long-term postoperative complications. Nine patients underwent incontinent ileovesicostomy between 2000 and 2013 at a mean age of 10.3 years (range 1.4 to 15.5). Surgical indication was sequelae of neurogenic or nonneurogenic neurogenic bladder (such as infection or worsening hydronephrosis) in 5 patients, reversal of vesicostomy in 3 and closure of cloacal exstrophy in 1. All 9 patients were thought incapable of reliable clean intermittent catheterization due to family unwillingness, poor social support or patient refusal. Median followup was 11.5 months (mean 48.2, range 1.3 to 144.8). Immediate postoperative complications included ileus requiring total parenteral nutrition and a wound infection in 1 patient. Long-term complications included urinary tract infection in 2 patients (febrile in 1 and positive culture for foul smelling urine in 1), stomal issues in 2 and temporary urethral leakage in 1. Constipation affected 3 children in long-term followup (all with neurogenic bowel preoperatively). Postoperative creatinine was stable or improved in all patients. Ileovesicostomy is a viable approach in children left with few other options, particularly those who are noncompliant or physically/socially unable to handle catheterization. This operation can help keep such patients out of diapers.


The rebound mechanism in catalytic C-H oxidation by MnO(tpp)Cl from DFT studies: electronic nature of the active species.
DFT studies show that the rebound mechanism for MnO(tpp)(Cl)-catalyzed C-H hydroxylation is favored for spin states with oxyl character.


A new formula to improve the screening of impaired glucose tolerance in youths with overweight or obesity.
To assess a new formula to improve the screening of isolated impaired glucose tolerance (IGT) in youth with overweight/obesity (OW/OB). A cross-sectional study was performed in 1189 Caucasian youths with OW/OB aged 5-17 years, in whom impaired fasting glucose and high glycosylated hemoglobin were excluded. The sample was divided into training set (TS) (n = 883) and validation set (VS) (n = 306). Fasting (FG) and post-load plasma glucose, alanine aminotransferase (ALT), lipids and familial history for type 2 diabetes (FD) were available in all individuals. In the TS youths with IGT (n = 58, 7.0%) showed higher prevalence of female sex (FS), FD, and higher levels of FG, post-load glucose, ALT and lower levels of HDL-cholesterol vs individuals without IGT. The linear formula was obtained by logistic regression analysis in the TS: 0.05∗ALT + 0.07∗FG + 0.87∗FD + (0.06∗HDL∗ - 1) + 1∗FS. The best cut-off was 5.84. The performance of the formula vs IGT was: sensitivity: 0.74 and specificity: 0.71. Similar results were obtained in the VS. Using metabolic and anamnestic data we obtained a simple formula with a good performance for screening isolated IGT. This formula may support pediatricians to identify youths with OW/OB in whom the OGTT may be useful for detecting IGT.


The role of hepatic microenvironment in hepatic fibrosis development.
Fibrosis is a common pathological feature of most types of chronic liver injuries. There is no specific treatment for liver fibrosis at present. The liver microenvironment, which fosters the survival and activity of liver cells, plays an important role in maintaining the normal structure and physiological function of the liver. The aim of this review is to deeply understand the role of the liver microenvironment in the dynamic and complicated development of liver fibrosis. After searching in Elsevier ScienceDirect, PubMed and Web of Science databases using 'liver fibrosis' and 'microenvironment' as keywords, studies related to microenvironment in liver fibrosis was compiled and examined. The homeostasis of the liver microenvironment is disrupted during the development of liver fibrosis, affecting liver cell function, causing various types of cell reactions, and changing the cell-cell and cell-matrix interactions, eventually affecting fibrosis formation. Liver microenvironment may be important for identifying potential therapeutic targets, and restoring microenvironment homeostasis may be an important strategy for promoting the reversal of liver fibrosis.KEY MESSAGESThe homeostasis of the liver microenvironment is disrupted in liver fibrosis;A pro-fibrotic microenvironment is formed during the development of liver fibrosis;Restoring microenvironment homeostasis may be an important strategy for promoting the reversal of liver fibrosis.


Improved Prognostic Performance of Right Atrial Pressure-Corrected Cardiac Power Output in Pulmonary Hypertension and Heart Failure with Preserved Ejection Fraction.
Cardiac power output (CPO) is a powerful predictor of adverse outcomes in heart failure (HF). However, the original formula of CPO included the difference between mean arterial pressure and right atrial pressure (RAP). The prognostic performance of RAP-corrected CPO (CPORAP) remains unknown in heart failure with preserved ejection fraction (HFpEF). We studied 101 HF patients with a left ventricular ejection fraction > 40% who had pulmonary hypertension due to left heart disease. CPORAP was significantly more discriminating than CPO in predicting outcomes (Delong test, P = 0.004). Twenty-five (24.8%) patients presented with dis-concordantly high CPORAP and low CPO when stratified by the identified CPORAP threshold of 0.547 W and the accepted CPO threshold of 0.803 W. These patients had the lowest RAP, and their cumulative incidence was comparable with those with concordantly high CPO and CPORAP (P = 0.313). CPORAP might identify patients with right ventricular involvement, thereby providing better prognostic performance than CPO in HFpEF.


Analysis and health risk assessment of toxic and essential elements of the wild fish caught by anglers in Liuzhou as a large industrial city of China.
Most wild fish caught by anglers (WFAs) are likely to be contaminated by toxic metals, particularly the fish collected from the waterways in urban and suburban areas; hence, the determination of health risk caused by WFAs consumption associated with toxic metals is vital. Therefore, Liuzhou, one of the largest industrial cities in China, was considered as an example city in this study. Eight toxic elements were analysed to uncover the pollution status and consumption safety of WFAs. Moreover, the suitable angling waterways were identified in the urban and suburban areas. The obtained results suggested relatively high concentrations of Zn, Cr and Cd, which were also found to be beyond corresponding Maximum Residue Limit. Among all analysed elements, only the mean pollution indices of Cr and Cd were observed to be beyond 0.2, revealing that the observed WFAs were generally contaminated by these metals. However, the potential health risk of WFAs can be predominately attributed to Cr, confirmed by the significantly higher Target hazard quotients (THQ). For adults, all the THQ values were below 1, indicating no significant health risk being associated with WFAs consumption in the case of adults. On the contrary, all the THQ values for Children were beyond 1, suggesting children being susceptible to great health risks due to WFAs consumption. Furthermore, the weekly recommended consumption of WFAs in urban area is remarkably lower than the current rate of fish consumption observed among urban residents; therefore, the waterways in urban areas can be evaded for fish angling.


Deciphering the DNA Damage Response.
This year's Albert Lasker Basic Medical Research Award honors Evelyn Witkin and Stephen J. Elledge, two pioneers in elucidating the DNA damage response, whose contributions span more than 40 years.


Synthesis, applications and biosafety evaluation of carbon dots derived from herbal medicine.
Carbon dots (CDs) are novel zero-dimensional spherical nanoparticles with water solubility, biocompatibility and photoluminescence properties. As the variety of raw materials for CDs synthesis becomes more and more abundant, people tend to choose precursors from nature. Many recent studies have shown that CDs can inherit properties similar to their carbon sources. Chinese herbal medicine has a variety of therapeutic effects to many diseases. In recent years, many literatures have chosen herbal medicine as raw materials, however, how the properties of raw materials affect CDs has not been systematically summarized. The intrinsic bioactivity and potential pharmacological effects of CDs have not received sufficient attention and have become a 'blind spot' for research. In this paper, the main synthesis methods were introduced and the effects of carbon sources from different herbal medicine on the properties of CDs and related applications were reviewed. In addition, we briefly review some of the biosafety assessments of CDs, and make recommendations for biomedical applications. CDs that inherit the therapeutic properties of herbs can enable diagnosis and treatment of clinical diseases, bioimaging, and biosensing in the future.


Parallel multigrid method for solving inverse problems.
We considered in this work the linear operator equation and used the Landweber iterative method as an iterative solver. After that, we used the multigrid method as an optimization method for obtaining an approximation solution with a highly accurate and fast process. A new parallel algorithm for the multigrid process has been developed. The proposed algorithm is based on a V-cycle mixed with the two-grid method. This modification of the V-cycle provides for parallel computing for each level. A coarse grid operator with a residual right-hand side vector for each coarse grid is provided. This parallel algorithm is used to accelerate and enhance computation for the solution of the iteration method in solving the inverse ill-posed problems. The necessary cost-time computation for all stages and processes for the parallel V-cycle algorithm has been done. A numerical experiment on solving the IVP (initial value problem) for the heat equation showed that the new parallel algorithm is much more efficient than the sequential method.•The study of iteration algorithms and mathematical experiments reveals a slow rate of convergence.•The Multigrid method is often used to speed up the rate of convergence of iterative methods, which is an effective method of solving large systems of linear algebra equations.•The approximation solution for the linear algebra equations was found by using the parallel method with the multigrid method.


Synthesis and conformational analysis of salivary proline-rich peptide P-B.
The 57-amino acid human salivary polypeptide P-B has been synthesized by the solid-phase method using 9-fluorenylmethoxycarbonyl (Fmoc) strategy. The circular dichroism (CD) spectroscopy, Fourier-transform infrared spectroscopy (FTIR) and molecular modeling methods have been used for conformational studies of P-B. Examination of the CD spectra of P-B showed the content of the secondary structure to be independent of temperature over the range 0-60 °C at pH = 7 as well as over the pH range of 2-12 at 37 °C. P-B adopts predominantly unordered structure with locally appearing β-turns. The cumulative results obtained using the CD and FTIR spectroscopic techniques indicate the percentage of the polyproline type-II (PPII) helix being as low as about 10%. Similarly, the molecular dynamics (MD) simulations reveal only a short PPII helix in the C-terminal fragment of the peptide (Pro(51)-Pro(54)), which constitutes 7%.


Effect of soy protein isolate nanoparticles loaded with litsea cubeba essential oil on performance of lentinan edible films.
Environmental issues caused by plastic packaging materials have gotten increasingly severe, and substantial research has been conducted on environmentally friendly active packaging materials. In this study, the Litsea cubeba essential oil loaded soy protein isolate nanoparticles (LSNPs) with appropriate particle size, high storage stability and salt solution stability were fabricated. The LSNPs with the highest encapsulation efficiency of 81.76 % were added into the lentinan edible film. The microstructures of the films were observed by scanning electron microscopy. The physical properties of the films were measured. The results show that the lentinan film with LSNPs in the volume ratio of 4:1 (LF-4) had the highest elongation at break of 196 %, the lowest oxygen permeability of 12 meq/kg, and good tensile strength, water vapor barrier property, antibacterial property, oxidation resistance and thermal stability. The study suggested that LF-4 film could inhibit the growth of bacteria and delay the oxidation of lipid and protein on beef surface for 7 d.


Free gracilis muscle transfer with ulnar nerve neurotization for elbow flexion restoration.
In upper and chronic brachial plexus injuries for which neurological surgery is not a good treatment option, one possibility for gaining elbow flexion is free functional muscle transfer. The primary aim of our study was to evaluate the elbow flexion gain achieved by free gracilis muscle transfer with partial ulnar nerve neurotization. This surgery was performed in 21 patients with upper and chronic (> 12 months) brachial plexus injuries. The level of injury, patient age, the time between trauma and surgery, the affected side, and the aetiology of the lesion were recorded. The primary outcome evaluated was elbow flexion muscle strength, which was measured using the British Medical Research Council (BMRC) scale, in patients with a minimum follow-up period of 12 months. The criterion used to classify elbow flexion as good was a grade of M4 or higher. An M4 elbow flexion strength gain was observed in 61.9% of the patients. A gain of M2 or higher was observed in 95.2% of the patients. The mean range of active motion was 77° (range 10 minimum-110 maximum). In patients with upper and chronic brachial plexus injuries, free gracilis muscle transfer with ulnar nerve neurotization yields a satisfactory gain in elbow flexion strength and is therefore a good treatment option.


Determinants of incident atherosclerotic cardiovascular disease events among individuals with type 2 diabetic microvascular complications in the UK: a prospective cohort study.
To evaluate the association of atherosclerotic cardiovascular disease (ASCVD) risk factors with incident ASCVD events among type 2 diabetes (T2D) individuals with microvascular complications. We included T2D participants with only microvascular complications from the UK Biobank cohort at baseline (2006-2010). Multivariable-adjusted Cox proportional hazards models were used to study the association between ASCVD risk factors with adjudicated incident ASCVD in T2D participants with only microvascular complications. A restricted cubic spline approach was employed to evaluate potential nonlinear associations between ASCVD risk factors and ASCVD. We studied 4,129 T2D individuals with microvascular complications at baseline. Over a median follow-up of 11.7 years, a total of 1,180 cases of incident ASCVD were documented, of which 1,040 were CHD, 100 were stroke, and 40 were both CHD and stroke events. After multivariable-adjustment, high-density lipoprotein cholesterol (HDL-C) level was linearly associated with a decreased risk of incident ASCVD [hazard ratio (HR): 0.49, 95% Confidence interval (CI): 0.32-0.75, Plinear = 0.011] and each 10 nmol/L increase of lipoprotein(a) [Lp(a)] level (HR: 1.02, 95% CI: 1.00-1.04, Plinear = 0.012) was linearly associated with an increased risk of incident ASCVD in T2D participants with only microvascular complications. HDL-C levels and Lp(a) levels (per 10 nmol/L) showed an independent linear relation with ASCVD risk among T2D individuals with only microvascular complications at long-term follow-up.


Forecasting changes in preference over the life span: a qualitative study of African-American men's prostate cancer decision making.
The goal of this study was to explore the processes by which African-American men, at greatest risk, might forecast and manage health changes as they age if they were diagnosed with prostate cancer. Twenty-nine African-American men, 40-70 years old, with no history of prostate cancer, participated in four focus groups and four follow-up individual interviews. Transcripts were analyzed using Grounded Theory, with thematic analysis and constant comparison of data. There was a curvilinear relationship between age and participants' preference for quality versus quantity of life in deciding to treat prostate cancer. Two mechanisms accounted for this: a change with age in the (1) reference point for judging value and (2) decision-making goal. With increasing long-term survivorship, it is vital to understand the multiple decisions cancer patients will face as they continue to age. The current study is an initial step in studying how patients might anticipate and manage such changes.


Social fragmentation, severe mental illness and suicide.
Geographic patterns of suicide are associated with area levels of social fragmentation. It is unknown whether this reflects higher levels of severe mental illness in socially fragmented areas. Data on psychiatric inpatient admissions and suicides amongst people aged 15-64 living in the City of Bristol [1991-1992] were postcode matched to the city's 34 electoral wards. Ecological associations of psychiatric admission (used as a 'proxy' measure of prevalence of severe mental illness) and suicide rates with levels of social fragmentation were investigated using negative binomial regression models. Psychiatric hospital admission rates were higher in areas with high levels of socioeconomic deprivation than in areas with high levels of social fragmentation. In contrast, associations with suicide were stronger in relation to social fragmentation than socioeconomic deprivation. Association of suicide with social fragmentation was only moderately attenuated in models controlling for psychiatric admission rate and socio-economic deprivation, RR 1.23 (95 % C. I. 1.09-1.38) per quartile increase in social fragmentation, compared to 1.29 (95% C. I. 1.16-1.44) before adjustment. The association between social fragmentation and suicide is not explained by socioeconomic deprivation or the prevalence of severe mental illness within socially fragmented areas as measured by psychiatric admission rate.


[Tumor-stroma ratio is an independent prognostic factor of non-small cell lung cancer].
The different expression of tumor-stroma ratio (TSR) have been proved to be a new and reliable independent prognostic factor in some solid tumors.The aim of the study is to test the different expression of TSR and its prognostic significance in non-small cell lung cancer (NSCLC). A total of 73 patients who underwent surgery resection for NSCLC were included in this study. TSR was assessed visually on the hematoxylin-stained tissue sections of surgical specimens. Patients with more than 50% intratumor stroma were quantified as the stroma-rich group and those with less than 50% as the stroma-poor group. In 73 cases of tissue samples, 46 cases were included in the stroma-poor group, while 27 cases in stroma-rich group. The different expression of TSR in NSCLC tissue was not correlated with gender, age and pathological type, lymph node metastasis, tumor size, pTNM staging, and so on. Kaplan-Meier survival analysis showed that the different expression of TSR was significantly correlated with survival days (P=0.014). Cox regression analysis showed that not only different expression of TSR is a independent prognostic factor for NSCLC (HR=1.832, 95%CI: 1.017-3.299), but also pTNM staging (HR=1.953, 95%CI: 1.284-2.970). The different expression of TSR might be an independent prognostic factor in NSCLC.


Modeling and simulations of the pharmacokinetics of fluorophore conjugated antibodies in tumor vicinity for the optimization of fluorescence-based optical imaging.
One of the methods to detect and localize tumors in tissue is to use fluorophore conjugated specific antibodies as tumor surface markers. The goals of this study are to understand and quantify the pharmacokinetics of fluorophore conjugated antibodies in the vicinity of a tumor. This study concludes another stage of the development of a non-invasive fluorescenated antibody-based technique for imaging and localization of tumors in vivo. A mathematical model of the pharmacokinetics of fluorophore conjugated antibodies in the vicinity of a tumor was developed based on histological staining experiments. We present the model equations of concentrations of antibodies and free binding sites. We also present a powerful simulation tool that we developed to simulate the imaging process. We analyzed the model and studied the effects of various independent parameters on the imaging result. These parameters included initial volume of markers (injected volume), total number of binding sites, tumor size, binding and dissociation rate constants, and the diffusion coefficient. We present the relations needed between these parameters in order to optimize the imaging results. A powerful and accurate tool was developed which may assist in optimizing the imaging system results by setting the injection volume and concentration of fluorophore conjugated antibodies in tissue and approximating the time interval where maximum specific binding occurs and the tumor can be imaged.


A retrospective study of endodontically treated mandibular first molars in a Chinese population.
The purpose of this investigation was to study the prevalence of three roots and four root canals in the mandibular first molars of a Chinese population. A total of 832 cases of mandibular first molars that had received root canal treatment clinically were examined by reviewing the clinical records and radiographs. The results showed that: (a) Mandibular first molars with four root canals occur at a frequency of 31.5%. (b) More than two-thirds (68.3%) of the above cases had the fourth canal arising from the third root, an extradistal root. (c) The bilateral occurrence of this extradistal root in mandibular first molars was 67%. In conclusion, the occurrence of four root canals in mandibular first molars among Chinese people was not unusual.


Clinical practice guidelines and performance measures in ESRD.
There is considerable variation in process or outcome measures in dialysis centers across the United States. Continuous quality improvement (CQI) techniques help multidisciplinary care teams assess current care and design strategies to improve outcomes. Evidence-based medicine facilitates the use of the current best evidence from clinical care research to improve patient care. Clinical practice guidelines, which are systematic statements that assist practitioner and patient decisions, are helpful in framing strategies to improve care. Clinical performance measures will provide measurement tools to CQI teams. Clinical decisions depend not only on research evidence in the literature, but also on clinical expertise, patient preferences, and constraints, such as policies, community standards, and budgetary limitations. As care teams use practice guidelines to improve outcomes, results of the CQI process may allow for the refinement of practice guidelines.


Tuning the Triplet Excited State of Bis(dipyrrin) Zinc(II) Complexes: Symmetry Breaking Charge Transfer Architecture with Exceptionally Long Lived Triplet State for Upconversion.
Zinc(II) bis(dipyrrin) complexes, which feature intense visible absorption and efficient symmetry breaking charge transfer (SBCT) are outstanding candidates for photovoltaics but their short lived triplet states limit applications in several areas. Herein we demonstrate that triplet excited state dynamics of bis(dipyrrin) complexes can be efficiently tuned by attaching electron donating aryl moieties at the 5,5'-position of the complexes. For the first time, a long lived triplet excited state (τT =296 μs) along with efficient ISC ability (ΦΔ =71 %) was observed for zinc(II) bis(dipyrrin) complexes, formed via SBCT. The results revealed that molecular geometry and energy gap between the charge transfer (CT) state and triplet energy levels strongly control the triplet excited state properties of the complexes. An efficient triplet-triplet annihilation upconversion system was devised for the first time using a SBCT architecture as triplet photosensitizer, reaching a high upconversion quantum yield of 6.2 %. Our findings provide a blueprint for the development of triplet photosensitizers based on earth abundant metal complexes with long lived triplet state for revolutionary photochemical applications.


Effect of abnormal pulmonary flow distribution on ventilatory efficiency and exercise capacity after arterial switch operation for transposition of great arteries.
Patients with anatomic repair of transposition of the great arteries (TGA) can present with branch pulmonary artery (PA) stenosis; however, its relation to an abnormal cardiopulmonary response to exercise is unknown. We investigated the relation between the PA anatomy and pulmonary blood flow (PBF) distribution and the cardiopulmonary response to exercise in patients with anatomic repair of TGA. We used cardiopulmonary exercise testing and magnetic resonance imaging to study 55 consecutive patients (62% male; age 14.4 ± 2.3 years) who had undergone neonatal anatomic repair of TGA. The peak oxygen uptake and slope of carbon dioxide elimination/minute ventilation was 79 ± 15% of predicted and 29.8 ± 3.8, respectively. Abnormal peak oxygen uptake (R = 0.363, p = 0.0082) and slope of carbon dioxide elimination/minute ventilation (R = 0.612, p <0.0001) values were associated with an abnormal right/left PBF distribution. However, although an increased ventilatory response to exercise appeared to be primarily related to an abnormal right/left PBF distribution, exercise capacity appeared to be related to the extent of the proximal PA branches and main PA stenosis (R = 0.476, p = 0.0004), suggesting that mechanical obstruction to PBF during exercise could be the main mechanism causing an abnormal exercise capacity. In conclusion, an abnormal PBF distribution related to branch PA stenosis or hypoplasia was associated with a reduced exercise capacity and increased ventilatory drive during exercise in patients with anatomic repair of TGA. Cardiopulmonary exercise test data can complement the anatomic and magnetic resonance imaging data in selecting those lesions that are functionally important.


Electromyography variables during the golf swing: a literature review.
The aim of the study was to review systematically the literature available on electromyographic (EMG) variables of the golf swing. From the 19 studies found, a high variety of EMG methodologies were reported. With respect to EMG intensity, the right erector spinae seems to be highly activated, especially during the acceleration phase, whereas the oblique abdominal muscles showed moderate to low levels of activation. The pectoralis major, subscapularis and latissimus dorsi muscles of both sides showed their peak activity during the acceleration phase. High muscle activity was found in the forearm muscles, especially in the wrist flexor muscles demonstrating activity levels above the maximal voluntary contraction. In the lower limb higher muscle activity of the trail side was found. There is no consensus on the influence of the golf club used on the neuromuscular patterns described. Furthermore, there is a lack of studies on average golf players, since most studies were executed on professional or low handicap golfers. Further EMG studies are needed, especially on lower limb muscles, to describe golf swing muscle activation patterns and to evaluate timing parameters to characterize neuromuscular patterns responsible for an efficient movement with lowest risk for injury.


Late onset MELAS with m.3243A > G mutation and its association with aneurysm formation.
We reported a 53-year-old with late-onset mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) accompanied by aneurysm and large vessel dilations. Most studies have focused on microangiopathy causing stroke-like episodes. We report a case to describe large vessel involvement in clinical considerations, and possible mechanisms of aneurysm formation. We recommended regular angiographic examination for patients with MELAS.


Risk factors of hip joint structural changes following pedicle subtraction osteotomy for ankylosing spondylitis-related thoracolumbar kyphosis: a minimum two-year observation.
To determine the clinical- and surgical-related factors of hip joint structural changes in ankylosing spondylitis (AS) patients with thoracolumbar kyphosis who underwent PSO. Hip involvement was assessed by the Bath Ankylosing Spondylitis Radiology Hip Index (BASRI-h) and defined by a score of at least 2. 52 patients with BASRI-h scores maintained and 78 patients with BASRI-h scores increased during follow-up were retrospectively reviewed. Clinical data were recorded. Radiological assessment was performed preoperatively, postoperatively, and at the final follow-up. No significant difference existed in age, gender and follow-up time between the groups, but earlier onset of AS, longer disease and kyphotic duration, and worse Bath Ankylosing Spondylitis Functional Index (BASFI) scores at the final follow-up were observed in patients with BASRI-h scores increased (P < 0.05). Besides, global kyphosis (GK), T1-pelvic angle (TPA), pelvic tilt (PT) and anterior pelvic plane angle (APPA) were always larger in patients with BASRI-h scores increased, accompanied with more sacral fixation (P < 0.05). Multivariate logistics regression showed that earlier onset of AS, longer kyphotic duration, larger preoperative GK, sacral fixation and larger APPA during follow-up were independent risk factors. Earlier onset of AS and longer kyphotic duration were the clinical risk factors of hip joint structural changes in AS patients following PSO, while larger preoperative GK, sacral fixation in PSO and larger APPA during follow-up were the surgical-related factors. Surgeons should inform patients with risk factors of the possibility for severe hip joint structural changes after PSO.


Vacuolar H(+)-ATPase-an enzyme for all seasons.
The life of every eukaryotic cell depends on the function of vacuolar H(+)-ATPase (V-ATPase). Because of its complexity and its challenging properties, the study of this enzyme has lagged behind that of its close relative, F-ATPase. We now know that V-ATPase is vital for many more physiological and biochemical processes than anticipated when the enzyme was discovered a few decades ago. It plays a crucial role in the proper functioning of internal organelles such as vacuoles, lysosomes, synaptic vesicles, endosomes, secretory granules, and the Golgi apparatus as well as in plasma membrane of several organisms and specific tissues and specialized cells. Knowledge of its involvement in several diseases, including cancer, has helped to establish the importance of V-ATPase for the preservation of life.


Effects of viscoelastic properties of tendon structures on stretch-shortening cycle exercise in vivo.
The aim of the present study was to examine the effects of viscoelastic properties of human tendon structures during stretch - shortening cycle exercise. The elongation of tendon and aponeurosis of the medial gastrocnemius muscle of 26 participants was measured by ultrasonography while they performed ramp isometric plantar flexion up to the voluntary maximum, followed by a ramp relaxation. The relationship between estimated muscle force and tendon elongation during the ascending phase was fitted to a linear regression, the slope of which was defined as stiffness. The percentage of the area within the muscle force-tendon elongation loop relative to the area beneath the curve during the ascending phase was defined as hysteresis. In addition, maximal voluntary concentric contractions at 2.09 and 3.14 rad x s(-1) with and without prior eccentric contractions were performed. The difference in the concentric torque at equivalent joint angles with and without prior eccentric contractions (i.e. pre-stretch augmentation) was negatively correlated with stiffness (P < 0.05) and hysteresis (P < 0.05). Furthermore, there was a higher correlation between the pre-stretch augmentation and the viscoelastic properties index--that is, the sum of normalized score values of stiffness and hysteresis (P < 0.01)--than with either stiffness or hysteresis alone. The results of this study suggest that performance during stretch-shortening cycle exercise is significantly affected by the viscoelastic properties of the tendon structures.


Two new lanostane-type triterpenoids from the fungus Periconia sp. TJ403-rc01.
The endophytic fungus Periconia sp. TJ403-rc01 (Dematiaceae) isolated from the leaves of Rosa chinensis Jacq. (Rosaceae) was cultivated on rice medium and chemically investigated, affording two new lanostane-type triterpenoids, namely pericinones A and B (1 and 2). Their structures were determined mainly by 1 D and 2 D NMR and HRESIMS data. Notably, it is the first report of lanostane-type triterpenoids from species of Periconia. Compounds 1 and 2 showed moderate anti-inflammatory activity against the NO production with IC50 values of 24.12 ± 0.73 and 11.38 ± 1.56 μM, respectively.


RPS: a comprehensive database of RNAs involved in liquid-liquid phase separation.
Liquid-liquid phase separation (LLPS) is critical for assembling membraneless organelles (MLOs) such as nucleoli, P-bodies, and stress granules, which are involved in various physiological processes and pathological conditions. While the critical role of RNA in the formation and the maintenance of MLOs is increasingly appreciated, there is still a lack of specific resources for LLPS-related RNAs. Here, we presented RPS (http://rps.renlab.org), a comprehensive database of LLPS-related RNAs in 20 distinct biomolecular condensates from eukaryotes and viruses. Currently, RPS contains 21,613 LLPS-related RNAs with three different evidence types, including 'Reviewed', 'High-throughput' and 'Predicted'. RPS provides extensive annotations of LLPS-associated RNA properties, including sequence features, RNA structures, RNA-protein/RNA-RNA interactions, and RNA modifications. Moreover, RPS also provides comprehensive disease annotations to help users to explore the relationship between LLPS and disease. The user-friendly web interface of RPS allows users to access the data efficiently. In summary, we believe that RPS will serve as a valuable platform to study the role of RNA in LLPS and further improve our understanding of the biological functions of LLPS.


An Integrative Approach to Elucidate Mechanisms Underlying the Pharmacokinetic Goldenseal-Midazolam Interaction: Application of In Vitro Assays and Physiologically Based Pharmacokinetic Models to Understand Clinical Observations.
The natural product goldenseal is a clinical inhibitor of CYP3A activity, as evidenced by a 40%-60% increase in midazolam area under the plasma concentration versus time curve (AUC) after coadministration with goldenseal. The predominant goldenseal alkaloids berberine and (-)-β-hydrastine were previously identified as time-dependent CYP3A inhibitors using human liver microsomes. Whether these alkaloids contribute to the clinical interaction, as well as the primary anatomic site (hepatic vs. intestinal) and mode of CYP3A inhibition (reversible vs. time-dependent), remain uncharacterized. The objective of this study was to mechanistically assess the pharmacokinetic goldenseal-midazolam interaction using an integrated in vitro-in vivo-in silico approach. Using human intestinal microsomes, (-)-β-hydrastine was a more potent time-dependent inhibitor of midazolam 1'-hydroxylation than berberine (KI and kinact: 8.48 μM and 0.041 minutes-1, respectively, vs. >250 μM and ∼0.06 minutes-1, respectively). Both the AUC and Cmax of midazolam increased by 40%-60% after acute (single 3-g dose) and chronic (1 g thrice daily × 6 days) goldenseal administration to healthy adults. These increases, coupled with a modest or no increase (≤23%) in half-life, suggested that goldenseal primarily inhibited intestinal CYP3A. A physiologically based pharmacokinetic interaction model incorporating berberine and (-)-β-hydrastine successfully predicted the goldenseal-midazolam interaction to within 20% of that observed after both chronic and acute goldenseal administration. Simulations implicated (-)-β-hydrastine as the major alkaloid precipitating the interaction, primarily via time-dependent inhibition of intestinal CYP3A, after chronic and acute goldenseal exposure. Results highlight the potential interplay between time-dependent and reversible inhibition of intestinal CYP3A as the mechanism underlying natural product-drug interactions, even after acute exposure to the precipitant. SIGNIFICANCE STATEMENT: Natural products can alter the pharmacokinetics of an object drug, potentially resulting in increased off-target effects or decreased efficacy of the drug. The objective of this work was to evaluate fundamental mechanisms underlying the clinically observed goldenseal-midazolam interaction. Results support the use of an integrated approach involving established in vitro assays, clinical evaluation, and physiologically based pharmacokinetic modeling to elucidate the complex interplay between multiple phytoconstituents and various pharmacokinetic processes driving a drug interaction.


Seeing the countryside through medical eyes: social service reports in the making of a sickly nation.
This article explores sanitary reports sent by early generations of social service year medical students who wrote about their first encounters with rural diseases and the people who suffered them. By exploring what was reported we see how, instead of questioning the roots of rural illness, poverty, and the hunger that they witnessed, young doctors often unwittingly reinforced urban prejudices and concerns that blamed indigenous Mexicans for their own poverty and diseased status. Because sanitary reports were authored by medical students, they were often perceived as 'scientific' evidence of the living conditions and unhealthy choices of rural Mexicans. The author argues that as in the case of travel narratives, medical students' written assessments influenced how the rest of society came to understand rural Mexico.


The trapezoid release for scar contractures: early experience.
A description is given of the technique of trapezoid release of scar contractures. Some clinical applications are presented and the results in 18 patients with a total of 37 flaps and maximum follow-up of 6 years are reported. A brief discussion of this method's advantages over existing techniques is presented.


Electrophysiological studies with new CCK analogs: correlation with binding affinity on B-type receptors.
The electrophysiological effects of Boc-D-Asp-Tyr(SO3H)-Nle-D-Lys-Trp-Nle-Asp-Phe-NH2 (compound I) and Boc-gamma-D-Glu-Tyr(SO3H)-Nle-D-Lys-Trp-Nle-Asp-Phe-NH2 (compound II), two cyclic cholecystokinin analogs with high selectivity for CCK-B receptors, as well as the effects of the linear enzyme-resistant analog Boc-[Nle28,Nle31]-CCK7 (BDNL), were compared with those of CCK8 using extracellular recordings in rat hippocampal slices in vitro. Bath applications of the three synthetic compounds resulted in concentration-dependent and reversible increases in single-unit activity. Comparison of equieffective concentrations yielded the following potency rank order: BDNL greater than CCK8 greater than compound II greater than compound I. There was a close correlation (r = .96, slope = 0.98) between the excitatory activities of the analogs and their potencies in displacing radiolabelled CCK8 from CCK-B receptors on rat brain membranes.


Ghettos and enclaves in the cross-place realm: mapping socially bounded spaces across cities.
Since the early Chicago School, urban researchers have used residential proximity to assess contacts within and between racial and ethnic groups. This approach is increasingly limited. Diverse groups use email, social networking sites, instant messaging and mobile phones to communicate across urban zones and distant cities. These practices enable mutual support among far-flung family members and co-ethnics as they engage with an array of institutions throughout their day. Through interviews and observations that include women and men of diverse occupations, races and national origins, the author explores how and why cross-place enclosures of sociality and resources develop. Rather than framing the residential area as the locus of racial/ethnic concentration, the author focuses on cross-place concentrations in the technologically mediated workspace. This study enhances theorization of the structural negotiations, interpersonal pressures and group preferences that produce separate lifeworlds in globalizing cities.


Cyclic hydrostatic pressure promotes uroplakin expression in human urothelial cells through activation of ERK1/2 signaling.
To investigate the effect of cyclic hydrostatic pressure on the expression of uroplakins and the role of extracellular regulated protein kinases 1/2 (ERK1/2) in the hydrostatic pressure-induced uroplakin expression of human urothelial cells (UCs). Human UCs were seeded into a cell culture flask and subjected to cyclic hydrodynamic pressures for 24 h. Pressure parameters were set as follows: static, 100 cm H2O, 200 cm H2O and 300 cm H2O pressure. Real-time polymerase chain reaction (RT-PCR) and western blot were used to detect the expression of uroplakins. The role of the ERK1/2 was investigated using ERK1/2 inhibitor. Compared with the 0 cm H2O control group, 200 cm H2O hydrostatic pressure significantly increased the expression of uroplakins, however, 100 cm and 300 cm pressures could not promote uroplakin expression. Hence, ERK1/2 expression was also detected under 200 cm H2O hydrostatic pressure. Western blot showed that 200 cm H2O pressure promoted the expression of ERK1/2. ERK1/2 inhibitor decreased the pressure-induced ERK1/2 activivation and uroplakin expression. Cyclic hydrostatic pressure increases the expression of uroplakins via activating ERK1/2 signaling pathway in human UCs, and 200 cm H2O pressure may be an optimal stress parameter to promote the uroplakin expression.


Quantitative explanation for increased affinity shown by mixtures of monoclonal antibodies: importance of a circular complex.
Mixtures of some but not all monoclonal antibodies which bind to separate epitopes on human chorionic gonadotropin (hCG) show an increased affinity for the hormone. To find an explanation for the increase in affinity, we developed a mathematical model which predicts the quantities of intermediates formed when pairs of IgG1 mouse monoclonal antibodies having affinities of approximately 10(8) M-1 for hCG are mixed with the hormone. At low antibody concentrations (i.e. less than 1 nM or 0.15 micrograms/ml) analysis of possible antibody-hormone combinations, including linear and circular chains composed of less than 12 molecules of antibody and 12 molecules of hCG, suggests the increase in affinity is due to formation of a circular complex containing two molecules of antibody and two of hCG. Further, the model predicts that the circular complex will be the major species formed at antibody-antigen equivalence. This prediction is supported by experimental observations on the molecular weight of a new complex formed in the presence of hCG and the mixture of the monoclonal antibodies. In addition, based on experimental values of binding constants for individual antibodies to hCG, the model correctly quantifies the loss in complex observed in the presence of excess hCG antigen. At high antibody concentrations (i.e. greater than 10 nM or 1.5 micrograms/ml) the formation of linear chains of antibody hCG pairs becomes appreciable and contributes to the increase in apparent affinity of the mixture for hCG. These results suggest that the observed affinity of complex mixtures of antibody for antigens containing multiple epitopes calculated from Scatchard plots may not be related to the affinity or avidity of any of the antibody species for a given epitope.


MR imaging of liver abscesses; application of Gd-DTPA.
The potential utility of Gd-DTPA contrast enhancement of MR images in the evaluation of liver abscesses was assessed in rodents. Twelve rats with surgically implanted sterile liver abscesses were imaged at various stages of focal hepatic inflammation, 48 hours, 4 days, 7 days, 14 days and 21 days after lesion induction. Spin echo images, acquired before and repeatedly after intravenous injection of 0.2 mmol/kg Gd-DTPA, demonstrated improvement of the lesion-to-background contrast ranging from 2% to 40% depending on the stage of the disease. The enhancement pattern also varied with abscess evolution. Two, four and seven-day-old abscesses typically showed a ring enhancement, whereas two- and three-week-old abscesses presented largely homogeneously enhancing lesions. In the earlier lesions, contrast enhanced rim surrounding the low intensity center corresponded histologically to the formation of a capsule consisting of fibrous tissue and inflammatory cells. The center was necrotic. Data show that abscesses can be detected on images acquired with long repetition and echo times without injection of Gd-DTPA. The administration of Gd-DTPA, however, improved the lesion-to-background contrast and helped to define the abscess capsule evolution.


Fabric-based exocervical and endocervical biopsy in comparison with punch biopsy and sharp curettage.
Fabric-based endocervical and exocervical biopsy tools were compared with exocervical punch biopsy and sharp endocervical curettage. We compared the level of pain reported by the patient and the amount of bleeding rated by the physician relative to the type of biopsy used. This was a 2-center institutional review board-approved study of patients undergoing colposcopy. One center randomized patients to undergo either conventional (punch biopsy, sharp endocervical curettage) or fabric-based biopsy of the endocervix and exocervix, and a second center enrolled nonrandomized controls. The patient and physician subjectively rated bleeding (exocervical) and pain (endocervical and exocervical) from the biopsy procedure(s), which were compared using the nonparametric Wilcoxon test. Exocervical and endocervical biopsies from 55 patients were compared. Fabric-based biopsies were abundant full-thickness curettage and showed statistically significant less pain and bleeding than the conventional biopsy methods (p < .0001). Fabric-based biopsies are significantly less traumatic than conventional biopsies. The benefits of a less-invasive biopsy could increase the willingness to perform multiple biopsies, increase disease detection, and improve patient satisfaction, which could impact future gynecologic health-seeking behavior and compliance with colposcopy recall.


High throughput sequencing of small RNAs transcriptomes in two Crassostrea oysters identifies microRNAs involved in osmotic stress response.
Increasing evidence suggests that microRNAs post-transcriptionally regulate gene expression and are involved in responses to biotic and abiotic stress. However, the role of miRNAs involved in osmotic plasticity remains largely unknown in marine bivalves. In the present study, we performed low salinity challenge with two Crassostrea species (C. gigas and C. hongkongensis), and conducted high-throughput sequencing of four small RNA libraries constructed from the gill tissues. A total of 202 and 87 miRNAs were identified from C. gigas and C. hongkongensis, respectively. Six miRNAs in C. gigas and two in C. hongkongensis were differentially expressed in response to osmotic stress. The expression profiles of these eight miRNAs were validated by qRT-PCR. Based on GO enrichment and KEGG pathway analysis, genes associated with microtubule-based process and cellular component movement were enriched in both species. In addition, five miRNA-mRNA interaction pairs that showed opposite expression patterns were identified in the C. hongkongensis, Differential expression analysis identified the miRNAs that play important regulatory roles in response to low salinity stress, providing insights into molecular mechanisms that are essential for salinity tolerance in marine bivalves.


Treatment choices for newly diagnosed primary open angle and ocular hypertension patients.
Despite advances in our knowledge of the aetiology and pathophysiology of glaucoma, the sole proven, effective intervention for treating primary open-angle glaucoma (POAG) and ocular hypertension (OHT) remains lowering of intraocular pressure (IOP) to prevent further progression and visual loss. The purpose of this review is to evaluate the treatment choices available to newly diagnosed POAG and OHT patients. We review the existing literature on treatments currently available to newly diagnosed POAG and OHT patients and discuss their role in the treatment paradigm of POAG and OHT. We consider different factors that may be important when offering a choice of treatment to newly diagnosed POAG and OHT patients as well as describing new glaucoma treatments in development and future directions for treatment.


[Electron microscopic and histochemical study of the localization of adenylate cylase and acetylcholinesterase in the synapses of the rat cerebral cortex and basal ganglia].
Synapses of the brain cortex and basal ganglia of the rat were studied by means of electron histochemical reactions to adenylate cyclase and acetylcholinesterase. Three types of synapses, viz. cholinergic, adrenergic and mixed, were identified. Typification was carried out on the basis of presynaptic terminal vesicle characteristics and localization of the mentioned enzymes of the synapse receptor area.


Type B monoamine-oxidase-containing cells and fibers in the cat hypothalamus demonstrated by an improved enzyme histochemical method.
The present study, using a diaminobenzidine (DAB)-coupled peroxidation method, examined the distribution and morphological characteristics of neuronal structures containing type B monoamine oxidase (MAO-B) in the cat hypothalamus. Large and intensely stained, distinctive MAO-B-positive cells, multipolar and with long dendritic arbors, were principally distributed in the ventral hypothalamus extending from A7 to A12.5 of the Horsley-Clarke plane. These cells were located caudally in the ventral surface of the brain including the tuberomamillary nucleus (TM) and the region surrounding the mamillary nuclei. Rostrally, they were aggregated in the area surrounding the fornix, particularly in the lateral perifornical region, and dispersed in the anterior mamillary nucleus, lateral hypothalamic area (HLA), and the medial tip of the entopeduncular nucleus. The most rostral positive cell group was identified in a narrow space between the optic tract and the entopeduncular nucleus at the A12.5 level. In addition to these large cells, the present study disclosed the presence of "small" to "very small" MAO-B-positive cells in the area surrounding the mamillary recess and the lateral part of the caudal arcuate nucleus. Distinct MAO-B-stained fibers were identified in all regions of the hypothalamus. A large number of thick labeled fibers were observed in the ventral hypothalamus including the TM and premamillary nucleus and posterior and lateral hypothalamic areas. A dense network of MAO-B-positive terminal-like fibers was observed in the dorsomedial nucleus where very small labeled cells were scattered. Many intensely stained thick and straight fibers were seen running ventrolaterally in the anterior part of the HLA and in the narrow space between the entopeduncular nucleus and optic tract. In the area of the tuber cinereum and the ventral part of the HLA, there were many positive fibers cut transversely, possibly projecting to the more anterior parts of the brain such as the diagonal band of Broca or septal nuclei.


Naturally occurring carbonyl compounds are mutagens in Salmonella tester strain TA104.
Strains of Salmonella typhimurium that carry a nonsense mutation at the site of reversion detect a variety of naturally occurring and synthetic carbonyl compounds as direct-acting mutagens. TA104 is reverted efficiently by formaldehyde, alpha, beta-unsaturated aldehydes (enals), and dicarbonyl compounds, such as diacetyl and glutaraldehyde. This strain is much more sensitive to carbonyl mutagenesis than is TA100, a strain previously reported to detect aldehydes as mutagens, or any other characterized strains of Salmonella. Long-chain enals are very toxic to TA104, but addition of a reduced glutathione chase following an incubation period decreases this toxicity, thus enabling the detection of 4-hydroxy-pentenal, a homolog of the lipid peroxidation product, 4-hydroxy-nonenal, as a mutagen. This is the first report of the mutagenicity of a hydroxy-enal, a class of enals produced by lipid peroxidation. Testing conducted with strains that carry the nonsense mutation in different repair backgrounds indicates that the presence of pKM101 and the deletion of the uvrB gene facilitate the detection of enals and dicarbonyls, but not malondialdehyde, as mutagens. Since carbonyl compounds are widely distributed in foods, are generated during cellular metabolism, and are present in body fluids, they may make a significant contribution to the risk of human cancer.


Cognitive impairments and mood disruptions negatively impact instrumental activities of daily living performance in the first three months after a first stroke.
Cognition and mood play crucial roles in post-stroke recovery; however, the stroke literature is unclear as to how impairments in both domains influence performance of instrumental activities of daily living (IADL). (1) Evaluate the extent to which mood and cognition at two weeks post-stroke predict performance three months post-stroke. (2) Assess performance differences in patients with impairments in both cognition and mood to patients with impairments in either cognition or mood. Inpatients with a first-ever ischemic or hemorrhagic stroke were assessed at 2 weeks (n = 52) and at 3 months (n = 41) post-stroke. Patients completed a battery of neuropsychological tests, self-report measures and performance-based tests. Cognitive impairments and mood disruptions were assessed at 2 weeks and three months and IADL performance, as assessed by the Executive Function Performance Test, was evaluated at three months. Complete data from the 41 patients assessed at both time points were analyzed. Regression analysis showed that composite cognition and composite mood variables at two weeks post-stroke predicted 48% of the variance in IADL performance at three months (F3,37 = 12.04; adjusted R(2) = 0.48, P < 0.001). Statistically significant differences were found in performance scores for patients with a single impairment (M = 7.86, SD = 7.81) and for those with impairments in both mood and cognition (M = 19.2, SD = 13.2) (t(39) = - 3.41, P = 0.008). The results of this study suggest that cognitive and mood impairments at two weeks post-stroke are important predictors of performance in complex activities required for full independence at home and should be routinely assessed in stroke rehabilitation.


Trends in Medicare Billing Among Mohs Surgeons in the United States During 2012: A Retrospective Cohort Study.
It is important to understand variability in practice patterns of Mohs surgeons. To examine the practice patterns of physicians performing Mohs micrographic surgery (MMS) in the United States. This retrospective cohort study of the 2012 Medicare Physician and Other Supplier Public Use Files includes all physicians who billed Medicare for MMS. The authors found 2,067 physicians who billed Medicare for MMS in 2012. American College of Mohs Surgery (ACMS) members took a significantly higher average number of head and neck (H&N) and trunk layers compared with American Society for Mohs Surgery (ASMS) members and those with no membership (p < .001). Male surgeons, surgeons with more experience (21+ years out), surgeons in private practice, and those practicing in rural populations closed a significantly greater proportion of cases with flaps or grafts, as compared to females (p < .001), those with less experience (<21 years out) (p < .001), surgeons in academic practice (p = .004), and those practicing in urban or cluster populations (p < .001), respectively. There is significant variability in practices of Mohs surgeons in the United States.


A novel brain tumour model in zebrafish reveals the role of YAP activation in MAPK- and PI3K-induced malignant growth.
Somatic mutations activating MAPK and PI3K signalling play a pivotal role in both tumours and brain developmental disorders. We developed a zebrafish model of brain tumours based on somatic expression of oncogenes that activate MAPK and PI3K signalling in neural progenitor cells and found that HRASV12 was the most effective in inducing both heterotopia and invasive tumours. Tumours, but not heterotopias, require persistent activation of phospho (p)-ERK and express a gene signature similar to the mesenchymal glioblastoma subtype, with a strong YAP component. Application of an eight-gene signature to human brain tumours establishes that YAP activation distinguishes between mesenchymal glioblastoma and low grade glioma in a wide The Cancer Genome Atlas (TCGA) sample set including gliomas and glioblastomas (GBMs). This suggests that the activation of YAP might be an important event in brain tumour development, promoting malignant versus benign brain lesions. Indeed, co-expression of dominant-active YAP (YAPS5A) and HRASV12 abolishes the development of heterotopias and leads to the sole development of aggressive tumours. Thus, we have developed a model proving that neurodevelopmental disorders and brain tumours might originate from the same activation of oncogenes through somatic mutations, and established that YAP activation is a hallmark of malignant brain tumours.


Inhibitory and excitatory synaptic neuroadaptations in the diazepam tolerant brain.
Benzodiazepine (BZ) drugs treat seizures, anxiety, insomnia, and alcohol withdrawal by potentiating γ2 subunit containing GABA type A receptors (GABAARs). BZ clinical use is hampered by tolerance and withdrawal symptoms including heightened seizure susceptibility, panic, and sleep disturbances. Here, we investigated inhibitory GABAergic and excitatory glutamatergic plasticity in mice tolerant to benzodiazepine sedation. Repeated diazepam (DZP) treatment diminished sedative effects and decreased DZP potentiation of GABAAR synaptic currents without impacting overall synaptic inhibition. While DZP did not alter γ2-GABAAR subunit composition, there was a redistribution of extrasynaptic GABAARs to synapses, resulting in higher levels of synaptic BZ-insensitive α4-containing GABAARs and a concomitant reduction in tonic inhibition. Conversely, excitatory glutamatergic synaptic transmission was increased, and NMDAR subunits were upregulated at synaptic and total protein levels. Quantitative proteomics further revealed cortex neuroadaptations of key pro-excitatory mediators and synaptic plasticity pathways highlighted by Ca2+/calmodulin-dependent protein kinase II (CAMKII), MAPK, and PKC signaling. Thus, reduced inhibitory GABAergic tone and elevated glutamatergic neurotransmission contribute to disrupted excitation/inhibition balance and reduced BZ therapeutic power with benzodiazepine tolerance.


Total joint arthroplasty versus resection-interposition arthroplasty for thumb carpometacarpal arthritis: a randomized controlled trial.
Thumb carpometacarpal (TCMC) osteoarthritis is a common condition that causes pain and functional limitations. We compared the outcomes of 2 surgical procedures for TCMC osteoarthritis, the Epping resection-suspension arthroplasty and the double-mobility TCMC prosthesis, and focused on pain relief, functional outcomes, and patient quality of life. Over a 7-year period a randomized controlled trial including 183 cases of TCMC osteoarthritis was conducted comparing a double mobility TCMC prosthesis (Moovis, Stryker, Kalamazoo, MI, USA) with the Epping resection-suspension arthroplasty. Pre- and postoperative examinations included the range of motion (ROM), SFMcGill score, visual analogue scale (VAS), the disabilities of the arm, shoulder and hand questionnaire (DASH), and the hospital anxiety and depression scale (HADS). At the 6-week postoperative follow-up, significant differences were found in VAS: Epping median 4.0 (interquartile range [IQR] 2.0-5.0) vs. TCMC prosthesis 2.0 (IQR 0.25-4.0), p = 0.03, effect size (area under the curve [AUC]) 0.64 (95% confidence interval [CI] 0.55-0.73), in DASH score: Epping 61 (IQR 43-75) vs. TCMC prosthesis 45 (IQR 29-57), p < 0.001, AUC 0.69 (CI 0.61- 0.78), and in radial abduction: Epping 55 (IQR 50-60) vs. TCMC prosthesis 62 (IQR 60-70), p = 0.001, AUC 0.70 (CI 0.61-0.79). No significant group differences were found at the 6- and 12-months follow-up. During the follow-up period, 3 of 82 prostheses had to be revised but there was no revision in the Epping group. The double mobility TCMC prosthesis had superior outcomes compared with the Epping procedure at 6 weeks; however, there were no significant differences in outcomes at 6 months and 1 year postoperatively. The implant survival rate of 96% after 12 months was acceptable.


Low pH-induced hemolysis of erythrocytes is related to the entry of the acid into cytosole and oxidative stress on cellular membranes.
HCl-induced lysis of mammalian erythrocytes, pretreated with DIDS, which is a specific inhibitor of the anion transport in their membranes, was markedly delayed. After acidification of a suspension of DIDS-inhibited cells, hemolysis was initiated by addition of a protonophore (Na-salicylate) at any moment chosen by will. These findings revealed that low-pH hemolysis depended on the rate of the transfer of acid equivalents into cytosole. Erythrocyte acid resistance was studied in a group of mammals and found to be inversely related to the rate of monovalent anion exchange in membranes which supported the above observations. In human erythrocytes, the critical level of cytosole acidification was found to be about pH 5.7 by measuring the acid equivalent absorbed by cells prior to hemolysis. HCl-induced hemolysis was also studied in human erythrocyte ghosts resealed with one-sixth of the initial hemoglobin content of cells. During the prelytic interval the ghosts suspended in isotonic NaCl/sucrose media shrunk, indicating an increase in ion permeability. The increase in prelytic permeability and hemolysis were strongly delayed in ghosts prepared from DIDS-treated cells, suggesting a uniform mechanism of lysing in cells and their ghosts. The prelytic increase in ion permeability was measured by the corresponding rate of ghost shrinkage and was found to be pH-dependent, with a high value below pH 3.4 and a very low one above pH 4.0. Compared to cells, the prelytic barrier impairment in ghosts had more mild character although it required greater concentration of cytosolic H+. While finally complete, hemolysis of cells was strongly delayed in the presence of catalase (500-1500 U/ml) and superoxide dismutase (200-600 U/ml) in hemolytic media. In conclusion, the acid-induced hemolysis could be associated with an oxidative injury of membranes, mainly triggered by the entry of acid equivalents into the cytosole.


The effects of percutaneous nephrolithotomy on renal function in geriatric patients in the early postoperative period.
Since percutaneous nephrolithotomy (PNL) is a surgical intervention, it may show deteriorative effects on renal function in the early postoperative period. In this study, the deteriorative effects of PNL on renal function were investigated in the geriatric population, and it was compared to the non-geriatric population.A total of 711 patients [64 in the geriatric group (female/male ratio: 33/31, mean age: 70.4 +/- 4.2 years), 647 in the non-geriatric group (female/male ratio: 267/380, mean age: 40.9 +/- 12.7 years)] were included in the study. Demographic data, biochemical parameters, stone diameters, duration of the operation and estimated glomerular filtration rates (eGFR) were recorded both preoperatively and postoperatively.The eGFR ratio (post-/preoperative) was 1.13 +/- 1.00 in the geriatric group, 0.98 +/- 0.20 in the non-geriatric group (P < 0.001). Although the mean stone diameter was larger in the geriatric group than the non-geriatric group (10.08 +/- 6.5 mm, 8.28 +/- 5.54 mm, P = 0.037), the mean duration of the operation was shorter in the geriatric group than in the non-geriatric group (55.7 +/- 13.6 min, 61.3 +/- 20.9 min, P = 0.036). Hematocrit, hemoglobin, urea levels, and grade of hydronephrosis were not different between the groups.Renal function impairment is expected to be common in the geriatric patient population since they already have lower basal renal function. In the present study, the ratio of postoperative to preoperative eGFR was higher in the geriatric group. Hematocrit, hemoglobin, and urea levels and degree of hydronephrosis of these two groups did not show a great difference. The relatively short duration of the operation in the geriatric group and the diversity of perioperative and postoperative care of these two groups may explain the difference.


Implementation of a Virtual Interprofessional ICU Learning Collaborative: Successes, Challenges, and Initial Reactions From the Structured Team-Based Optimal Patient-Centered Care for Virus COVID-19 Collaborators.
Initial Society of Critical Care Medicine Discovery Viral Infection and Respiratory illness Universal Study (VIRUS) Registry analysis suggested that improvements in critical care processes offered the greatest modifiable opportunity to improve critically ill COVID-19 patient outcomes. The Structured Team-based Optimal Patient-Centered Care for Virus COVID-19 ICU Collaborative was created to identify and speed implementation of best evidence based COVID-19 practices. This 6-month project included volunteer interprofessional teams from VIRUS Registry sites, who received online training on the Checklist for Early Recognition and Treatment of Acute Illness and iNjury approach, a structured and systematic method for delivering evidence based critical care. Collaborators participated in weekly 1-hour videoconference sessions on high impact topics, monthly quality improvement (QI) coaching sessions, and received extensive additional resources for asynchronous learning. Outcomes included learner engagement, satisfaction, and number of QI projects initiated by participating teams. Eleven of 13 initial sites participated in the Collaborative from March 2, 2021, to September 29, 2021. A total of 67 learners participated in the Collaborative, including 23 nurses, 22 physicians, 10 pharmacists, nine respiratory therapists, and three nonclinicians. Site attendance among the 11 sites in the 25 videoconference sessions ranged between 82% and 100%, with three sites providing at least one team member for 100% of sessions. The majority reported that topics matched their scope of practice (69%) and would highly recommend the program to colleagues (77%). A total of nine QI projects were initiated across three clinical domains and focused on improving adherence to established critical care practice bundles, reducing nosocomial complications, and strengthening patient- and family-centered care in the ICU. Major factors impacting successful Collaborative engagement included an engaged interprofessional team; an established culture of engagement; opportunities to benchmark performance and accelerate institutional innovation, networking, and acclaim; and ready access to data that could be leveraged for QI purposes. Use of a virtual platform to establish a learning collaborative to accelerate the identification, dissemination, and implementation of critical care best practices for COVID-19 is feasible. Our experience offers important lessons for future collaborative efforts focused on improving ICU processes of care.


MgATP specifically controls in vitro self-assembly of vertebrate skeletal myosin in the physiological pH range.
The appearances in the electron microscope of rat and rabbit skeletal muscle myosin filaments and rod aggregates, formed in the presence of variable amounts of MgATP, were compared at different pH values. It is shown that small amounts of MgATP, similar to those sufficient to trigger the dissociation of the actomyosin complex, were able to modify the geometry of myosin filaments profoundly in the physiological pH range, whereas the conformation of rod aggregates remained unchanged even in the presence of high concentrations of MgATP. Myosin filaments formed in the absence of MgATP displayed the classical spindle-shaped conformation and variable diameters at all pH values, whereas myosin filaments formed in the presence of MgATP in the physiological pH range had constant diameters, similar to those of natural thick filaments. These filaments of constant diameter frayed, rapidly and reversibly, into two types of subfilaments with respective diameters of 4 to 5 nm and 9 to 10 nm, when the pH of the medium was raised above 7.2. Spindle-shaped myosin filaments and rod aggregates remained unchanged by such small changes in pH. It was possible to change the conformation of preformed spindle-shaped filaments by simply adding MgATP to the medium, but this reaction was slow and took several hours to be completed. Relatively high concentrations of MgATP, similar to those in the living cell, increased the solubility of both myosin filaments and rod aggregates in the alkaline pH range (pH greater than or equal to 7.0). Low pH values (less than or equal to 6.5) and excess free Mg2+ (greater than or equal to 6 to 7 mM) abolished both the specific effect of MgATP on myosin filament conformation and its solubilizing effect on both myosin filaments and rod aggregates. The degree of purity of the myosin preparations and the level of phosphorylation of the LC-2 light chains did not influence filament behaviour noticeably and rat and rabbit myosins behaved similarly.


The Global Impact of COVID-19 on Craniomaxillofacial Surgeons: A Follow-Up Survey After One Year.
Comparative cross-sectional. To measure the impact that COrona VIrus Disease-19 (COVID-19) has had on craniomaxillofacial (CMF) surgeons after 1 year and compare it with 2020 data by (1) measuring access to adequate personal protective equipment (PPE), (2) performance of elective surgery, and (3) the vaccination status. This should be investigated because most CMF surgeons felt that hospitals did not provide them with adequate PPE. The investigators surveyed the international AO CMF membership using a 30-item online questionnaire and compared it to a previous study. The primary predictor variable was year of survey administration. Primary outcome variables were availability of adequate personal protective equipment (adequate/inadequate), performance of elective surgery (yes/no), and vaccination status (fully vaccinated/partly vaccinated/not vaccinated). Descriptive and analytic statistics were computed. Binary logistic regression models were created to measure the association between year and PPE availability. Statistical significance level was set at P < .05. The sample was composed of 523 surgeons (2% response rate). Most surgeons reported access to adequate PPE (74.6%). The most adequate PPE was offered in Europe (87.8%) with the least offered in Africa (45.5%). Surgeons in 2021 were more likely to report adequate PPE compared to 2020 (OR 3.74, 95% CI [2.59-4.39]). Most of the respondents resumed elective surgery (79.5% vs 13.3% in 2020) and were fully vaccinated (59.1%). Most CMF surgeons now have access to adequate PPE, resumed elective surgery, and are either fully or partly vaccinated. Future studies should investigate the long-term impact of the fast-evolving COVID-19 pandemic on CMF surgeons.


The effects on the plankton community of filter-feeding Sacramento blackfish, Orthodon microlepidotus.
A series of mesocosm experiments was performed to assess the effects on the plankton community of filter-feeding Sacramento blackfish (Cyprinidae; Orthodon microlepidotus). Phytoplankton size-frequency distribution, zooplankton abundance, primary production, potential secondary production, and nutrient concentrations were measured. Blackfish reduce numbers of both evasive and nonevasive zooplankton and large phytoplankton while enhancing nanoplankton densities. Blackfish also significantly increase primary production and potential secondary community production. Levels of dissolved inorganic phosphorus and ammonia-nitrogen also increase. The effects of blackfish are generally similar to those reported for other filter-feeding fish.


Effect of ozone stress on crop productivity: A threat to food security.
Tropospheric ozone (O3), the most important phytotoxic air pollutant, can deteriorate crop quality and productivity. Notably, satellite and ground-level observations-based multimodel simulations demonstrate that the present and future predicted O3 exposures could threaten food security. Hence, the present study aims at reviewing the phytotoxicity caused by O3 pollution, which threatens the food security. The present review encompasses three major aspects; wherein the past and prevailing O3 concentrations in various regions were compiled at first, followed by discussing the physiological, biochemical and yield responses of economically important crop species, and considering the potential of O3 protectants to alleviate O3-induced phytotoxicity. Finally, the empirical data reported in the literature were quantitatively analysed to show that O3 causes detrimental effect on physiological traits, photosynthetic pigments, growth and yield attributes. The review on prevailing O3 concentrations over various regions, where economically important crop are grown, and their negative impact would support policy makers to implement air pollution regulations and the scientific community to develop countermeasures against O3 phytotoxicity for maintaining food security.


Pro-apoptotic activity of transiently expressed BCL-2 occurs independent of BAX and BAK.
BCL-2 suppresses apoptosis induced by a wide variety of stimuli in multiple cell types. Most of the in vitro studies that have examined the activity of BCL-2 have employed stable cell lines that ectopically express BCL-2. We have reported that BCL-2 is expressed at high levels in the absence of the 5'- and 3'-UTRs of the Bcl-2 gene and transient high level of expression results in potent cell death (Uhlmann et al., [1998]: JBC 278:17926-17932). Expression of BCL-2 under the transcriptional control of the cognate 5'- and 3'-UTRs express lower levels of BCL-2 and does not cause cell death. Our present results suggest that in contrast to BCL-2, transient expression of BCL-xL does not induce cell death and coexpression of BCL-xL with the pro-apoptotic BCL-2 does not suppress cell death. The pro-apoptotic activity of BCL-2 appears to involve activation of the cytochrome c/caspase 9/caspase 3 pathway. Elevated levels of BCL-2 expression results in N-terminal cleavage of BCL-2 at a novel site different from a previously identified caspase cleavage site at Asp 34 by a non-caspase protease. Transient expression of a BCL-2 mutant lacking aa 51-85 within the loop region induces efficient cell death and N-terminal cleavage of BCL-2 while a different deletion mutant lacking aa 30-91 induces reduced levels of cell death in the absence of BCL-2 cleavage suggesting that N-terminal processing of BCL-2 may be an amplification event in BCL-2-mediated cell death. Overexpression of BCL-2 in a Bax-null human colon cancer cell line (HCT116Bax-/-) induces efficient cell death. The pro-apoptotic activity of BCL-2 is also observed in a Bax-null cells in which BAK expression is inhibited by stable RNAi expression. Our results suggest that BCL-2 contains an intrinsic pro-apoptotic activity and can induce apoptosis independent of BAX and BAK under specific conditions.


Chemical sterilisation of Bos indicus bull calves following intratesticular injection of zinc acetate: Effects on growth and concentrations of testosterone.
The aim of this study was to determine the effects in Bos indicus calves of intra-testicular injection of either saline (n=9) or one of two doses of zinc acetate ((ZA1, 57.75mg, n=10, or ZA2, 71.75mg, n=10) or surgical castration (n=9) on circulating concentrations of testostosterone and liveweight. Human chorionic gonadotrophin (hCG, 1500IU) was administered 202 and 525 days after treatment on Day 0 and animals were slaughtered on Day 860. In animals left intact treatment with ZA reduced mean serum concentrations of testosterone (Saline: 5.58±0.79ng/mL, ZA1: 1.28±0.27ng/mL, ZA2: 1.01±0.17ng/mL; P<0.001) and concentrations 48h following administration of hCG. The maximum concentration of testosterone recorded throughout the study in six out of 19 animals treated with ZA was ≤0.21ng/mL. Treatment with ZA did not significantly affect live weights or carcass weights or result in any detectable scrotal lesions. Animals with concentrations of testosterone ≥1.0ng/mL exhibited greater liveweights throughout most of the study and yielded heavier carcass weights (340.9±7.02 versus 309.3±6.17kg, P=0.002). It is concluded that a single, intra-testicular administration of either 57.75mg or 71.75mg of ZA was able to similarly reduce circulating concentrations of testosterone without significantly affecting liveweights or carcass weights. Treatment with ZA can result in variation in circulating concentrations of testosterone which could lead to differences in behaviour, liveweights and carcass characteristics.


Isolation, culture and characterization of primary mouse RPE cells.
Mouse models are powerful tools for the study of ocular diseases. Alterations in the morphology and function of the retinal pigment epithelium (RPE) are common features shared by many ocular disorders. We report a detailed protocol to collect, seed, culture and characterize RPE cells from mice. We describe a reproducible method that we previously developed to collect and culture murine RPE cells on Transwells as functional polarized monolayers. The collection of RPE cells takes ∼3 h, and the cultures mimic in vivo RPE cell features within 1 week. This protocol also describes methods to characterize the cells on Transwells within 1-2 weeks by transmission and scanning electron microscopy (TEM and SEM, respectively), immunostaining of vibratome sections and flat mounts, and measurement of transepithelial electrical resistance. The RPE cell cultures are suitable to study the biology of the RPE from wild-type and genetically modified strains of mice between the ages of 10 d and 12 months. The RPE cells can also be manipulated to investigate molecular mechanisms underlying the RPE pathology in the numerous mouse models of ocular disorders. Furthermore, modeling the RPE pathology in vitro represents a new approach to testing drugs that will help accelerate the development of therapies for vision-threatening disorders such as macular degeneration (MD).