Specific Inhibition of Orai1-mediated Calcium Signalling Resolves Inflammation and Clears Bacteria in an Acute Respiratory Distress Syndrome Model.

Rationale: Acute respiratory distress syndrome (ARDS) has an unacceptably high mortality rate (35%) and is without effective therapy. Orai1 is a Ca2+ channel involved in store-operated Ca2+ entry (SOCE), a process that exquisitely regulates inflammation. Orai1 is considered a druggable target, but no Orai1-specific inhibitors exist to date. Objectives: To evaluate whether ELD607, a first-in-class Orai1 antagonist, can treat ARDS caused by bacterial pneumonia in preclinical models. Methods: ELD607 pharmacology was evaluated in HEK293T cells and freshly isolated immune cells from patients with ARDS. A murine acute lung injury model caused by bacterial pneumonia was then used: mice were infected with Pseudomonas aeruginosa, Staphylococcus aureus, methicillin-resistant S. aureus, or multidrug-resistant P. aeruginosa and then treated with ELD607 intranasally. Measurements and Main Results: ELD607 specifically inhibited SOCE in HEK293T cells with a half-maximal inhibitory concentration of 9 nM. ELD607 was stable in ARDS airway secretions and inhibited SOCE in ARDS immune cells. In vivo, inhaled ELD607 significantly reduced neutrophilia and improved survival. Surprisingly, Orai1 inhibition by ELD607 caused a significant reduction in lung bacteria, including methicillin-resistant S. aureus. ELD607 worked as an immunomodulator that reduced cytokine levels, reduced neutrophilia, and promoted macrophage-mediated resolution of inflammation and clearance of bacteria. Indeed, when alveolar macrophages were depleted with inhaled clodronate, ELD607 was no longer able to resolve inflammation or clear bacteria. Conclusions: These data indicate that specific Orai1 inhibition by ELD607 may be a novel approach to reduce multiorgan inflammation and treat antibiotic-resistant bacteria.

Partial dynamical symmetry versus quasi dynamical symmetry examination within a quantum chaos analyses of spectral data for even-even nuclei.

Statistical analyses of the spectral distributions of rotational bands in 51 deformed prolate even-even nuclei in the 152 ≤ A ≤ 250 mass region [Formula: see text] are examined in terms of nearest neighbor spacing distributions. Specifically, the focus is on data for 0+, 2+, and 4+ energy levels of the ground, gamma, and beta bands. The chaotic behavior of the gamma band, especially the position of the [Formula: see text] band-head compared to other levels and bands, is clear. The levels are analyzed within the framework of two models, namely, a SU(3)-partial dynamical symmetry Hamiltonian and a SU(3) two-coupled quasi-dynamical symmetry Hamiltonian, with results that are further analyzed using random matrix theory. The partial and quasi dynamics both yield outcomes that are in reasonable agreement with the known experimental results. However, due to the degeneracy of the beta and gamma bands within the simplest SU(3) picture, the theory cannot be used to describe the fluctuation properties of excited bands. By changing relative weights of the different terms in the partial and quasi dynamical Hamiltonians, results are obtained that show more GOE-like statistics in the partial dynamical formalism as the strength of the pairing term is increased. Also, in the quasi-dynamical symmetry limit, more correlations are found because of the stronger couplings.

Cigarillos Compromise the Mucosal Barrier and Protein Expression in Airway Epithelia.

Smoking remains a leading cause of preventable morbidity and mortality worldwide. Despite a downward trend in cigarette use, less-regulated tobacco products, such as cigarillos, which are often flavored to appeal to specific demographics, such as younger people, are becoming increasingly popular. Cigar/cigarillo smoking has been considered a safer alternative to cigarettes; however, the health risks associated with cigar in comparison with cigarette smoking are not well understood. To address this knowledge gap, we characterized the effects of multiple brands of cigarillos on the airway epithelium using ex vivo and in vivo models. To analyze these effects, we assessed the cellular viability and integrity of smoke-exposed primary airway cell cultures. We also investigated the protein compositions of apical secretions from cigarillo-exposed airway epithelial cultures and BAL fluid of cigarillo-exposed mice through label-free quantitative proteomics and determined the chemical composition of smoke collected from the investigated cigarillo products. We found that cigarillo smoke exerts similar or greater effects than cigarette smoke in terms of reduced cell viability; altered protein levels, including those of innate immune proteins; induced oxidative-stress markers; and greater nicotine delivery to cells. The analysis of the chemical composition of the investigated cigarillo products revealed differences that might be linked to the differential effects of these products on cell viability and protein abundance profiles, which have been associated with a range of health risks in the context of airway biology. These findings contradict the assumption that cigarillos might be safer and less harmful than cigarettes. Instead, our results indicate that cigarillo smoke is associated with equal or greater health risks and the same or increased airway toxicity compared with cigarette smoke.

Little Cigars are More Toxic than Cigarettes and Uniquely Change the Airway Gene and Protein Expression.

Little cigars (LCs) are regulated differently than cigarettes, allowing them to be potentially targeted at youth/young adults. We exposed human bronchial epithelial cultures (HBECs) to air or whole tobacco smoke from cigarettes vs. LCs. Chronic smoke exposure increased the number of dead cells, lactate dehydrogenase release, and interleukin-8 (IL-8) secretion and decreased apical cilia, cystic fibrosis transmembrane conductance regulator (CFTR) protein levels, and transepithelial resistance. These adverse effects were significantly greater in LC-exposed HBECs than cigarette exposed cultures. LC-exposure also elicited unique gene expression changes and altered the proteomic profiles of airway apical secretions compared to cigarette-exposed HBECs. Gas chromatography-mass spectrometry (GC-MS) analysis indicated that LCs produced more chemicals than cigarettes, suggesting that the increased chemical load of LCs may be the cause of the greater toxicity. This is the first study of the biological effects of LCs on pulmonary epithelia and our observations strongly suggest that LCs pose a more severe danger to human health than cigarettes.

Effects of acute or chronic ethanol exposure during adolescence on behavioral inhibition and efficiency in a modified water maze task.

Ethanol is well known to adversely affect frontal executive functioning, which continues to develop throughout adolescence and into young adulthood. This is also a developmental window in which ethanol is misused by a significant number of adolescents. We examined the effects of acute and chronic ethanol exposure during adolescence on behavioral inhibition and efficiency using a modified water maze task. During acquisition, rats were trained to find a stable visible platform onto which they could escape. During the test phase, the stable platform was converted to a visible floating platform (providing no escape) and a new hidden platform was added in the opposite quadrant. The hidden platform was the only means of escape during the test phase. In experiment 1, adolescent animals received ethanol (1.0 g/kg) 30 min before each session during the test phase. In experiment 2, adolescent animals received chronic intermittent ethanol (5.0 g/kg) for 16 days (PND30 To PND46) prior to any training in the maze. At PND72, training was initiated in the same modified water maze task. Results from experiment 1 indicated that acute ethanol promoted behavioral disinhibition and inefficiency. Experiment 2 showed that chronic intermittent ethanol during adolescence appeared to have no lasting effect on behavioral disinhibition or new spatial learning during adulthood. However, chronic ethanol did promote behavioral inefficiency. In summary, results indicate that ethanol-induced promotion of perseverative behavior may contribute to the many adverse behavioral sequelae of alcohol intoxication in adolescents and young adults. Moreover, the long-term effect of adolescent chronic ethanol exposure on behavioral efficiency is similar to that observed after chronic exposure in humans.

Effects of ethanol, Δ(9)-tetrahydrocannabinol, or their combination on object recognition memory and object preference in adolescent and adult male rats.

Recent advances have been made in our understanding of the deleterious effects of both ethanol and THC on adolescent behavior and brain development. However, very little is known about the combined effects of EtOH+THC during adolescence, a time in which these drugs are often used together. The purpose of this experiment was to: (1) determine whether EtOH and/or THC induced greater working memory impairment in adolescent than adult male rats using the novel object recognition (NOR) task and (2) determine whether the EtOH+THC combination would produce a more potent additive effect in adolescents than adults when compared to these drugs alone. NOR was performed with a 24h delay under each of the four drug conditions: vehicle; 1.5g/kg ethanol; 1.0mg/kg THC; and 1.5g/kg EtOH+1.0mg/kg THC, at 72h intervals. The results show that there was an age effect on working memory in NOR after the EtOH+THC challenge. Specifically, adolescent animals showed a preference for the familiar object whereas adults showed no preference for the novel or familiar object, the latter being characteristic of a classic working memory deficit. These effects were not dependent on changes in exploration across session, global activity across drug condition, or total object exploration. These novel findings clearly indicate that further understanding of this age-drug interaction is crucial to elucidating the influence that adolescent EtOH+THC use may have on repeated drug use and abuse later in life.

Outcome of living donor liver transplantation for Egyptian patients with hepatitis C (genotype 4)-related cirrhosis.

BACKGROUND: Hepatitis C virus (HCV) recurrence after living donor liver transplantation (LDLT) represents a challenging issue due to universal viral recurrence and invasion into the graft, although the incidence of histological recurrence, risk factors, and survival rates are still controversial. PATIENTS AND METHODS: Recurrence of HCV was studied in 38 of 53 adult patients who underwent LDLT. RESULTS: Recipient and graft survivals were 86.6% at the end of the follow-up which was comparable to literature reports for deceased donor liver transplantation (DDLT). Clinical HCV recurrence was observed in 10/38 patients (26.3%). Four patients developed mild fibrosis with a mean fibrosis score of 0.6 and mean grade of histological activity index (HAI) of 7.1. None of the recipients developed allograft cirrhosis during the mean follow-up period of 16 +/- 8.18 months (range, 4-35 months). Estimated and actual graft volumes were negatively correlated with the incidence and early clinical HCV recurrence. None of the other risk factors were significantly correlated with clinical HCV recurrence: gender, donor and recipient ages, pretransplantation Child-Pugh or model for end-stage liver disease (MELD) scores, pre- and postoperative viremia, immunosuppressive drugs, pulse steroid therapy, and preoperative anti-HBc status. CONCLUSIONS: Postoperative patient and graft survival rates for HCV (genotype 4)-related cirrhosis were more or less comparable to DDLT reported in the literature. Clinical HCV recurrence after LDLT in our study was low. Small graft volume was a significant risk factor for HCV recurrence. A longer follow-up and a larger number of patients are required to clarify these issues.

Effect of early feed restriction on reproductive performance in Japanese quail (Coturnix coturnix japonica).

Reproductive performance of quail (Coturnix coturnix japonica) was evaluated following feed restriction (100, 85, and 70% of ad libitum) between 2 to 5 wk of age with three replicates of 12, 2-wk-old chicks per replicate. Body weight, feed conversion, and leucocyte distribution were measured during feed restriction. After experimental feed treatment, BW, age at first egg, egg production, fertility, hatchability, and embryonic mortality were evaluated from five replicates of two females and one male per treatment. Feed-restricted female chicks had lower BW from 3 to 5 wk of age, but male weights were depressed only during the most severe restriction at 4 and 5 wk. No treatment differences were observed among BW within a sex from 6 to 13 wk. Body weights at first egg were significantly heavier for females fed 70% ad libitum than for birds on other treatments. Fertility, age at first egg, feed conversion, egg production, and egg weight were unaffected by feed restriction. Although hatchability was unaffected by feed restriction, percentage of late dead and total dead embryos were significantly reduced in eggs from restricted quail. Thirty quail fed 70% of ad libitum control intake had significantly increased egg specific gravity. Feed restriction increased the percentage of heterophils and basophils and the heterophil/lymphocyte ratio, whereas the percentage of lymphocytes and eosinophils decreased. Feed can be restricted to 85 or 70% of ad libitum feed intake from 2 to 5 wk of age without detrimentally affecting reproductive parameters between 6 to 13 wk of age.

Effect of acetyl salicylic acid in drinking water on reproductive performance of Japanese quail (Coturnix coturnix japonica).

The impact of acetyl salicylic add (ASA) on reproductive performance during periods of heat stress was studied in Japanese quail (Coturnix coturnix japonica). During a season when daily high temperatures averaged between 34 and 40 degrees C, four treatments each containing four replicates of five males and five females were given 0, 0.5, 1.0, or 1.5 g/L ASA in drinking water from 34 to 45 wk of age. At 39 wk of age control quail were lighter than quail fed ASA. Quail receiving 1.5 g/L ASA exhibited increased fertility, hatchability, and egg production and decreased late dead and total embryonic mortality. Early dead embryos, pips, chick weight, and egg weight were not different among treatments. The lumen of uterovaginal junction sperm host glands (UVJSHG) from control quail was apparently devoid of spermatozoa, whereas quail receiving 1.5 g/L ASA had the greatest apparent accumulation of spermatozoa. Control caudal infundibulum host glands (CPOI) were devoid of spermatozoa in contrast with CPOI from quail receiving ASA. Results show that UVJSHG lumens from quail receiving 1.5 g/L ASA had larger inside diameters than other treatment groups. However, the CPOI inside diameter and the outside diameters of both UVJSHG and CPOI were not affected by ASA. Adding 1.5 g/L ASA to the drinking water of quail under heat stress improved body weight, fertility, hatchability, embryonic mortality, egg production, egg specific gravity, and sperm storage capacity.

Effect of feeding time on the reproductive performance of Japanese quail (Coturnix coturnix japonica).

Several feeding regimens are applied to improve performance of fertile egg production during times of heat stress. During a period of heat stress (34 to 36 degrees C), two feeding periods were used to measure the impact of feeding time on reproductive performance of Japanese quail (Coturnix cotunix japonica) between 44 and 60 wk of age. Each feeding time treatment had 25 individually caged male and female pairs. Quail were fed ad libitum between 0600 to 1400 or 1400 to 2200 h daily. Results indicated that feeding between 0600 to 1400 h reduced BW, fertility, hatchability, egg production, and egg specific gravity when compared with the effects of feeding between 1400 to 2200 h. Feeding time had no effect on total embryonic mortality, egg weight, or the period between subsequent ovipositions. The different feeding times affected the distribution curve of oviposition over time. An instrument designed to record oviposition time is described. Results showed that selection of the time of day for application of an 8-h restricted feeding regimen affected BW, fertility, hatchability, egg production, egg specific gravity, and oviposition time in Japanese quail.

Seroepidemiological prevalence of hepatitis delta virus in eastern Saudi Arabia.

To describe hypercholesterolemia in an urban community in Saudi Arabia, total serum cholesterol (TSC) was measured in 966 apparently healthy males (475 Saudi Arabs, 351 other Arabs and 140 non-Arabs) using a portable analyzer (Boehringer Model[R]). Mean age was 35.9 years (SD+/-9.1) and mean body mass index (BMI) was 26.2 kg/m(2) (SD+/-3.4) with a mean TSC of 5.20 mmol (SD+/-1.21). Mean TSC was significantly higher among non-Arabs at 5.74 mmol/L (SD+/-1.48) than in Saudi Arabs at 4.93 mmol/L (SD+/-1.11), P<0.001 mmol/L. TSC was higher than 5.20 mmol/L in 44.3% and higher than 6.80 mmol/L in 6.9% of the population. TSC was higher than 6.80 mmol/L in 3.6% of Saudi Arabs, 8.0% in other Arabs and 15.7% in the non-Arabs. This calls for cholesterol screening of the indigenous male population for hypercholesterolemia and other coronary heart disease risk factors at every opportunity. Among expatriate males, a mass screening strategy might be appropriate. The health care system needs appropriate adjustment to deal with this growing health problem.

Measurements of genetic variation in residual metabolizable energy intake of laying hens.

An experiment was conducted to determine heritability of the residual component of feed consumption. The effect of test period length was also examined to determine the shortest test period that would result in reliable data. Three hundred and thirty-five pedigreed White Leghorn hens were used. The experiment began when the hens were 26 wk of age and lasted for 28 wk. The residual component of energy intake was determined for each hen for time periods of 2, 4, and 8 wk in length. These values were used to determine heritability. Analyses of the data for each period were performed in two steps. A reduced model, without pedigree information, was used to develop a regression model. A complete model was used for analyses of data. The procedure of starting egg mass output measurements 2 days after body weight measurements was more effective than starting records on the same days that body weights were obtained. The partial regression coefficients of ME intake decreased with age. Also the partial regression coefficients of ME on body weight change increased with the length of the test period. The partial regression coefficients of ME on egg mass increased slightly with age. Heritability of ME intake adjusted for metabolic body weight, body weight change, and egg mass output was highest (.567 +/- .186) when the test was calculated by 8-wk periods. However, the range of estimates over time was lowest when made for 4-wk periods.

Comparison of energy utilization efficiency among six lines of White Leghorns.

Individual records were used of feed consumption, BW, BW change, and egg mass (EM) output during a 4-wk period for 180 hens from six different lines of White Leghorn chickens (30 hens per line). The diet contained 16.45% protein and 2,825 kcal ME/kg. Feed consumption was used to calculate actual ME intake (AcME) for each hen. Multiple regression of AcME was performed with metabolic BW, BW change, and EM production as independent variables. The regression coefficients and the independent variables were then used to calculate the expected ME intake (ExME). The following values were calculated for each hen and were used to compare energy efficiency among the six lines: the residual component of energy intake (RES), defined as the difference between the AcME intake and ExME intake; the relative efficiency ratio (RER), defined as (ExME divided by AcME) X 100 (calculated using the partial regression coefficients of the overall equation); and feed conversion, the ratio of feed consumed to EM. Significant differences among the lines in efficiency of EM produced were observed regardless of the efficiency measurement used. Feed consumption did not indicate differences in RES or RER. A multiple regression equation was derived for each line. Significant differences were found among the six lines in their energy requirements for maintenance and for egg production. These results suggest the possible use of the partial regression coefficients as selection criteria for separately improving maintenance efficiency and egg production efficiency.