Comparison of Xrn1 and Rat1 5' → 3' exoribonucleases in budding yeast supports the specific role of Xrn1 in cotranslational mRNA decay.

The yeast Saccharomyces cerevisiae and most eukaryotes carry two 5' → 3' exoribonuclease paralogs. In yeast, they are called Xrn1, which shuttles between the nucleus and the cytoplasm, and executes major cytoplasmic messenger RNA (mRNA) decay, and Rat1, which carries a strong nuclear localization sequence (NLS) and localizes to the nucleus. Xrn1 is 30% identical to Rat1 but has an extra ~500 amino acids C-terminal extension. In the cytoplasm, Xrn1 can degrade decapped mRNAs during the last round of translation by ribosomes, a process referred to as "cotranslational mRNA decay." The division of labor between the two enzymes is still enigmatic and serves as a paradigm for the subfunctionalization of many other paralogs. Here we show that Rat1 is capable of functioning in cytoplasmic mRNA decay, provided that Rat1 remains cytoplasmic due to its NLS disruption (cRat1). This indicates that the physical segregation of the two paralogs plays roles in their specific functions. However, reversing segregation is not sufficient to fully complement the Xrn1 function. Specifically, cRat1 can partially restore the cell volume, mRNA stability, the proliferation rate, and 5' → 3' decay alterations that characterize xrn1Δ cells. Nevertheless, cotranslational decay is only slightly complemented by cRat1. The use of the AlphaFold prediction for cRat1 and its subsequent docking with the ribosome complex and the sequence conservation between cRat1 and Xrn1 suggest that the tight interaction with the ribosome observed for Xrn1 is not maintained in cRat1. Adding the Xrn1 C-terminal domain to Rat1 does not improve phenotypes, which indicates that lack of the C-terminal is not responsible for partial complementation. Overall, during evolution, it appears that the two paralogs have acquired specific characteristics to make functional partitioning beneficial.

Protective efficacy of Barbervax® in Merino weaner sheep trickle infected with five doses of Haemonchus contortus infective larvae.

Barbervax® protects sheep against H. contortus infection; however, the level of protection afforded by the vaccine at different levels of larval challenge under field conditions has not been reported. Therefore, this study was designed to investigate the protective efficacy of Barbervax® at increasing doses of trickle infection with H. contortus third stage larvae (L3). Merino lambs (220) were randomly allocated to challenge treatment groups and received Barbervax® vaccinations at approximately 8, 11, 15 and 22 weeks of age or not vaccinated (control group). Animals within each treatment group were subjected to one of five levels of H. contortus L3 trickle infection (0, 300, 600, 1200 and 2400 L3/week in two split doses). Trickle infections started two weeks after the third vaccination (week 0). Worm egg count (WEC), packed cell volume (PCV), antibody titre and bodyweight were measured at week 2 (start of trickle infection), week 5 and then every two weeks for 11 weeks. Vaccinated weaners had a significantly (P < 0.0001) lower faecal WEC than unvaccinated control animals. Vaccination induced significant specific antibody responses that were not influenced by level of trickle infection. Vaccination significantly reduced the number of weaners requiring salvage anthelmintic treatment (9.4% vaccinated weaners versus 33.6% unvaccinated). Vaccine protective index based on WEC was similar at all challenge levels (overall mean> 60%) and at the higher challenge levels did not reduce H. contortus infection to levels that would not require anthelmintic treatment. Therefore, it is suggested that under conditions of high larval challenge the use of Barbervax® should be integrated with other control methods.

The Emergence of a vv + MDV Can Break through the Protections Provided by the Current Vaccines.

Marek's disease (MD) is an infectious malignant T-cell lymphoma proliferative disease caused by Marek's disease virus (MDV). In recent years, the emergence of very virulent (vv) and/or very virulent plus (vv +) strains of MDV in the field has been suggested as one of the causes of vaccination failure. The pathogenicity of the MDV strain GX18NNM4, isolated from a clinical outbreak in a broiler breeder flock that was vaccinated with CVI988/Rispens, was investigated. In the vaccination-challenge test, GX18NNM4 was able to break through the protections provided by the vaccines CVI988 and 814. It also significantly reduced body weight gain and caused marked gross lesions and a large area of infiltration of neoplastic lymphocyte cells in the heart, liver, pancreas, etc. of the infected birds. In addition, the expressions of programmed death 1 (PD-1) and its ligand, programmed death ligand 1 (PD-L1), in the spleens and cecal tonsils (CTs) of the unvaccinated challenged birds were significantly increased compared to those in the vaccinated challenged birds, indicating that the PD-1/PD-L1 pathway is related to immune evasion mechanisms. The results showed that the GX18NNM4 strain could cause severe immunosuppression and significantly decrease the protections provided by the current commercial vaccines, thus showing GX18NNM4 to be a vv + MDV strain.

Comparative evaluation of antidotal efficacy of 2-PAM and HNK-102 oximes during inhalation of sarin vapor in Swiss albino mice.

Efficacy of two oximes treatments evaluated during inhalation of sarin vapor (LCt50, 755.9 mg/min/m3) in simulated real scenario in vivo. Majority of mice either became moribund or died within 1-2 min during exposure to multifold-lethal concentrations of sarin vapor. Protection indices were determined by exposing to sarin vapor in two sessions, 1 min exposure followed by treatments with or without HNK-102 (56.56 mg/kg, im) or 2-PAM (30 mg/kg, im) and atropine (10 mg/kg, ip), and again exposed for remaining 14 min. Protection offered by HNK-102 was found to be four folds higher compared to 2-PAM in the same toxic environment. Secondly, sub-lethal concentration of sarin vapor (0.8 × LCt50 or 605 mg/min/m3), 24 h post investigations revealed that the oximes could not reactivate brain and serum acetylcholinesterase (AChE) activity. The treatments prevented increase in protein concentration (p < .05) and macrophages infiltration compared to sarin alone group in broncho-alveolar lavage fluid. Lung histopathology showed intense peribronchial infiltration and edema with desquamating epithelial lining and mild to moderate alveolar septal infiltration in sarin and atropine groups, respectively. Noticeable peeling-off observed in epithelial lining and sporadic mild infiltration of epithelial cells at bronchiolar region in 2-PAM and HNK-102 groups, respectively. The oximes failed to reactivate AChE activity; however, the mice survived up to 6.0 × LCt50, proved involvement of non-AChE targets in sarin toxicity. Atropine alone treatment was found to be either ineffective or increased the toxicity. HNK-102, exhibited better survivability with lung protection, can be considered as a better replacement for 2-PAM to treat sarin inhalation induced poisoning.

Effect of health protective factors on health deficit accumulation and mortality risk in older adults in the Beijing Longitudinal Study of Aging.

OBJECTIVES: To evaluate transitions in health status and risk of death in older adults in relation to baseline health deficits and protective factors. DESIGN: Prospective cohort study with reassessments at 5, 8, and 15 years. SETTING: Secondary analysis of data from the Beijing Longitudinal Study on Aging. PARTICIPANTS: Urban and rural community-dwelling people aged 55 and older at baseline (n = 3,275), followed from 1992 to 2007, during which time 51% died. MEASUREMENTS: Health status was quantified using the deficit accumulation-based frailty index (FI), constructed from 30 intrinsic health measures. A protection index was constructed using 14 extrinsic items (e.g., exercise, education). The probabilities of health changes, including death, were evaluated using a multistate transition model. RESULTS: Women had more health deficits (mean baseline FI 0.13 ± 0.11) than did men (mean baseline FI 0.11 ± 0.10). Although health declined on average (mean FIs increased), improvement and stability were common. Baseline health significantly affected health transitions and survival over various follow-up durations (odds ratio (OR) = 1.27, 95% confidence interval (CI) = 1.17-1.37 for men; OR = 1.24, 95% CI = 1.16-1.33 for women for each increment of deficits). Each protective factor reduced the risk of health decline and the risk of death in men and women by 13% to 25%. CONCLUSION: Deficit accumulation-based transition modeling demonstrates persisting effects of baseline health status on age-related health outcomes. Some mitigation by protective factors can be demonstrated, suggesting that improving physical and social conditions might be beneficial.

Determinación de la influencia de la altitud en el índice de protección natural (IPN) de la piel y el factor de protección solar requerido (SPF) / Determination of the influence of altitude on the contents of natural protection (IPN) skin and sun protection factor required (SPF)

Se evalúa la influencia de la altitud sobre el Índice de Protección Natural (IPN) y el Factor de Protección Solar requerido (SPF) comparando niños de Mucuchies, Mérida, Venezuela (3100 m.s.n.m.) y Palmarito, Mérida, Venezuela (5 m.s.n.m.), utilizando el equipo Sun Protection Diagnostic SP 37®. Según el análisis de los resultados a través de estadísticas descriptivas, se evidencia que el IPN y el SPF son variables dependientes de la altitud de la localidad en que se encuentran los niños, ya que en las localidades más altas los rayos solares inciden de forma más directa que en las más bajas.

It is evaluated the effect of altitude on Natural Protection Index (IPN) and the required Sun Protection Factor (SPF) comparing children Mucuchies, Merida, Venezuela (3100 m) and Palmarito, Merida, Venezuela (5 m) using Sun Protection Diagnostic SP37® equipment. According to the analysis of results through descriptive statistics, it appears that the IPN and the SPF are variable dependent on the altitude of the locality in which children are, as in the most high solar rays more directly than in the lowest.

A single-center study evaluating the effect of the controlled adverse environment (CAE(SM)) model on tear film stability.

PURPOSE: To investigate use of an improved ocular tear film analysis protocol (OPI 2.0) in the Controlled Adverse Environment (CAE(SM)) model of dry eye disease, and to examine the utility of new metrics in the identification of subpopulations of dry eye patients. METHODS: Thirty-three dry eye subjects completed a single-center, single-visit, pilot CAE study. The primary endpoint was mean break-up area (MBA) as assessed by the OPI 2.0 system. Secondary endpoints included corneal fluorescein staining, tear film break-up time, and OPI 2.0 system measurements. Subjects were also asked to rate their ocular discomfort throughout the CAE. Dry eye endpoints were measured at baseline, immediately following a 90-minute CAE exposure, and again 30 minutes after exposure. RESULTS: The post-CAE measurements of MBA showed a statistically significant decrease from the baseline measurements. The decrease was relatively specific to those patients with moderate to severe dry eye, as measured by baseline MBA. Secondary endpoints including palpebral fissure size, corneal staining, and redness, also showed significant changes when pre- and post-CAE measurements were compared. A correlation analysis identified specific associations between MBA, blink rate, and palpebral fissure size. Comparison of MBA responses allowed us to identify subpopulations of subjects who exhibited different compensatory mechanisms in response to CAE challenge. Of note, none of the measures of tear film break-up time showed statistically significant changes or correlations in pre-, versus post-CAE measures. CONCLUSION: This pilot study confirms that the tear film metric MBA can detect changes in the ocular surface induced by a CAE, and that these changes are correlated with other, established measures of dry eye disease. The observed decrease in MBA following CAE exposure demonstrates that compensatory mechanisms are initiated during the CAE exposure, and that this compensation may provide the means to identify and characterize clinically relevant subpopulations of dry eye patients.

Validation and verification of the OPI 2.0 System.

PURPOSE: The Ocular Protection Index (OPI) 2.0 System was developed to evaluate ocular surface protection under a natural blink pattern and normal visual conditions. The OPI 2.0 System implements fully automated software algorithms which provide a real-time measurement of corneal exposure (breakup area) for each interblink interval during a 1-minute video. Utilizing this method, the mean breakup area (MBA) and OPI 2.0 (MBA/interblink interval) were calculated and analyzed. The purpose of this study was to verify and validate the OPI 2.0 System for its ability to distinguish between dry eye and normal subjects, and to accurately identify breakup area. METHODS: In order to verify and validate the OPI 2.0 System, a series of artificial images and a series of still image frames captured during an actual clinical session using fluorescein staining videography were analyzed. Finally, a clinical validation process was completed to determine the effectiveness and clinical relevance of the OPI 2.0 System to differentiate between dry eye and normal subjects. RESULTS: Software analysis verification conducted in a set of artificially constructed images and in actual videos both saw minimal error rates. MBA and OPI 2.0 calculations were able to distinguish between the qualifying eyes of dry eye and normal subjects in a statistically significant fashion (P < 0.001 for both outcomes). As expected, dry eye subjects had a higher MBA and OPI 2.0 than normal subjects (0.232, dry eye; 0.040, normal and 0.039, dry eye; 0.006, normal, respectively). Results for the worst eyes and all qualifying analyses based on staining, forced-stare tear film breakup time, and MBA were numerically similar. CONCLUSION: The OPI 2.0 System accurately identifies the degree of breakup area on the cornea and represents an efficient, clinically relevant measurement of the pathophysiology of the ocular surface.

Measurement of ocular surface protection under natural blink conditions.

PURPOSE: To evaluate a new method of measuring ocular exposure in the context of a natural blink pattern through analysis of the variables tear film breakup time (TFBUT), interblink interval (IBI), and tear film breakup area (BUA). METHODS: The traditional methodology (Forced-Stare [FS]) measures TFBUT and IBI separately. TFBUT is measured under forced-stare conditions by an examiner using a stopwatch, while IBI is measured as the subject watches television. The new methodology (video capture manual analysis [VCMA]) involves retrospective analysis of video data of fluorescein-stained eyes taken through a slit lamp while the subject watches television, and provides TFBUT and BUA for each IBI during the 1-minute video under natural blink conditions. The FS and VCMA methods were directly compared in the same set of dry-eye subjects. The VCMA method was evaluated for the ability to discriminate between dry-eye subjects and normal subjects. The VCMA method was further evaluated in the dry eye subjects for the ability to detect a treatment effect before, and 10 minutes after, bilateral instillation of an artificial tear solution. RESULTS: Ten normal subjects and 17 dry-eye subjects were studied. In the dry-eye subjects, the two methods differed with respect to mean TFBUTs (5.82 seconds, FS; 3.98 seconds, VCMA; P = 0.002). The FS variables alone (TFBUT, IBI) were not able to successfully distinguish between the dry-eye and normal subjects, whereas the additional VCMA variables, both derived and observed (BUA, BUA/IBI, breakup rate), were able to successfully distinguish between the dry-eye and normal subjects in a statistically significant fashion. TFBUT (P = 0.034) and BUA/IBI (P = 0.001) were able to distinguish the treatment effect of artificial tears in dry-eye subjects. CONCLUSION: The VCMA methodology provides a clinically relevant analysis of tear film stability measured in the context of a natural blink pattern.

Ocular iontophoresis of EGP-437 (dexamethasone phosphate) in dry eye patients: results of a randomized clinical trial.

PURPOSE: To assess safety and efficacy of EGP-437 (dexamethasone phosphate 40 mg/mL [DP]) in dry eye patients. METHODS: The study employed a prospective, single-center, double-masked design utilizing a Controlled Adverse Environment (CAE). Patients (n = 103) with confirmed signs and symptoms of dry eye syndrome were randomized into 1 of 3 iontophoresis treatment groups: 7.5 mA-min at 2.5 mA (DP 7.5, n = 41); 10.5 mA-min at 3.5 mA (DP 10.5, n = 37); or 10.5 mA-min at 3.5 mA (placebo, n = 25). Three CAE visits and 4 follow-up visits occurred over 3 weeks. Patients meeting enrollment criteria received iontophoresis in both eyes after the second CAE exposure (visit 3) and before the third CAE exposure (visit 5). Primary efficacy endpoints were corneal staining and ocular discomfort. Secondary endpoints included tear film break-up time, ocular protection index (OPI), and symptomatology. RESULTS: The DP 7.5 and DP 10.5 treatment groups showed statistically significant improvements in signs and symptoms of dry eye at various time points; however, the primary endpoints were not achieved. The DP 7.5 treatment group exhibited statistically significant improvements in corneal staining (when comparing the differences between study entry and exit, 3 weeks, P = 0.039), OPI (immediately following the second treatment, P = 0.048) and ocular discomfort at follow-up visits (a week after the first treatment, P = 0.032; 24 hours after the second treatment, P = 0.0032). Treatment-emergent adverse events (AEs) were experienced by 87% of patients and were consistent across all treatment groups. Most AEs were mild and no severe AEs were observed. CONCLUSION: Ocular iontophoresis of EGP-437 demonstrated statistically and clinically significant improvements in signs and symptoms of dry eye syndrome within a CAE model.