CEP Biomarkers as Potential Tools for Monitoring Therapeutics

Background

Carboxyethylpyrrole (CEP) adducts are oxidative modifications derived from docosahexaenoate-containing lipids that are elevated in ocular tissues and plasma in age-related macular degeneration (AMD) and in rodents exposed to intense light. The goal of this study was to determine whether light-induced CEP adducts and autoantibodies are modulated by pretreatment with AL-8309A under conditions that prevent photo-oxidative damage of rat retina. AL-8309A is a serotonin 5-HT_1A receptor agonist.

Methods

Albino rats were dark adapted prior to blue light exposure. Control rats were maintained in normal cyclic light. Rats were injected subcutaneously 3x with 10 mg/kg AL-8309A (2 days, 1 day and 0 hours) before light exposure for 6 h (3.1 mW/cm², λ=450 nm). Animals were sacrificed immediately following light exposure and eyes, retinas and plasma were collected. CEP adducts and autoantibodies were quantified by Western analysis or ELISA.

Results

ANOVA supported significant differences in mean amounts of CEP adducts and autoantibodies among the light + vehicle, light + drug and dark control groups from both retina and plasma. Light-induced CEP adducts in retina were reduced ~20% following pretreatment with AL-8309A (n = 62 rats, p = 0.006) and retinal CEP immunoreactivity was less intense by immunohistochemistry. Plasma levels of light-induced CEP adducts were reduced at least 30% (n = 15 rats, p = 0.004) by drug pretreatment. Following drug treatment, average CEP autoantibody titer in light exposed rats (n = 22) was unchanged from dark control levels, and ~20% (p = 0.046) lower than in vehicle-treated rats.

Conclusions

Light-induced CEP adducts in rat retina and plasma were significantly decreased by pretreatment with AL-8309A. These results are consistent with and extend previous studies showing AL-8309A reduces light-induced retinal lesions in rats and support CEP biomarkers as possible tools for monitoring the efficacy of select therapeutics.     

Phage Display and Synthetic Peptides as Promising Biotechnological Tools for the Serological Diagnosis of Leprosy

Background

The diagnosis of leprosy is primarily based on clinical manifestations, and there is no widely available laboratory test for the early detection of this disease, which is caused by Mycobacterium leprae. In fact, early detection and treatment are the key elements to the successful control of leprosy.

Methodology/Principal Findings

Peptide ligands for antibodies from leprosy patients were selected from phage-displayed peptide libraries. Three peptide sequences expressed by reactive phage clones were chemically synthesized. Serological assays that used synthetic peptides were evaluated using serum samples from leprosy patients, household contacts (HC) of leprosy patients, tuberculosis patients and endemic controls (EC). A pool of three peptides identified 73.9% (17/23) of multibacillary (MB) leprosy patients using an enzyme-linked immunosorbent assay (ELISA). These peptides also showed some seroreactivities to the HC and EC individuals. The peptides were not reactive to rabbit polyclonal antisera against the different environmental mycobacteria. The same peptides that were conjugated to the carrier protein bovine serum albumin (BSA) induced the production of antibodies in the mice. The anti-peptide antibodies that were used in the Western blotting analysis of M. leprae crude extracts revealed a single band of approximately 30 kDa in one-dimensional electrophoresis and four 30 kDa isoforms in the two-dimensional gel. The Western blotting data indicated that the three peptides are derived from the same bacterial protein.

Conclusions/Significance

These new antigens may be useful in the diagnosis of MB leprosy patients. Their potentials as diagnostic reagents must be more extensively evaluated in future studies using a large panel of positive and negative sera. Furthermore, other test approaches using peptides should be assessed to increase their sensitivity and specificity in detecting leprosy patients. We have revealed evidence in support of phage-displayed peptides as promising biotechnological tools for the design of leprosy diagnostic serological assays.     

Geographic Tools for Eradication Programs of Insular Non-native Mammals

Non-native mammals are major drivers of ecosystem change and biodiversity loss; this is especially apparent on islands. However, techniques exist to remove non-native mammals, providing a powerful conservation tool. Conservation practitioners are now targeting larger islands for restoration. Leveraging existing and developing new techniques and technologies will prove critical to successful eradications on large islands. Using the removal of introduced goats (Capra hircus) from Santiago Island, Galápagos as a case study, we present a suite of Geographic Information System (GIS) tools that aid island conservation actions. GIS tools were incorporated into the three phases of the eradication campaign: planning, hunting, and monitoring. Further, these tools were adopted for three eradication techniques: ground-based hunting, aerial hunting by helicopter, and Judas goats. These geographic approaches provide a foundation for statistical, spatial, and economic analyses that should increase the capability and efficiency of removal campaigns. Given limited conservation funds and the dire status of many insular species, efficiently removing non-native mammals from islands is of paramount global conservation importance.     

Monitoring of Mass Distribution Interventions for Trachoma in Plateau State,Nigeria

Mass drug administration (MDA) with antibiotics is a key component of the SAFE strategy for trachoma control. Guidelines recommend that where MDA is warranted the whole population be targeted with 80% considered the minimum acceptable coverage. In other countries, MDA is usually conducted by salaried Ministry of Health personnel (MOH). In Plateau State, Nigeria, the existing network of volunteer Community Directed Distributors (CDD) was used for the first trachoma MDA. We conducted a population-based cluster random survey (CRS) of MDA participation to determine the true coverage and compared this to coverage reported from CDD registers. We surveyed 1,791 people from 352 randomly selected households in 24 clusters in three districts in Plateau State in January 2011, following the implementation of MDA. Households were enumerated and all individuals present were asked about MDA participation. Household heads were questioned about household-level characteristics and predictors of participation. Individual responses were compared with the CDD registers. MDA coverage was estimated as 60.3% (95% CI 47.9–73.8%) by the survey compared with 75.8% from administrative program reports. CDD registration books for comparison with responses were available in 19 of the 24 clusters; there was a match for 658/682 (96%) of verifiable responses. CDD registers did not list 481 (41.3%) of the individuals surveyed. Gender and age were not associated with individual participation. Overall MDA coverage was lower than the minimum 80% target. The observed discrepancy between the administrative coverage estimate from program reports and the CRS was largely due to identification of communities missed by the MDA and not reported in the registers. CRS for evaluation of MDA provides a useful additional monitoring tool to CDD registers. These data support modification of distributor training and MDA delivery to increase coverage in subsequent rounds of MDA.     

An Assessment of the Effectiveness of High Definition Cameras as Remote Monitoring Tools for Dolphin Ecology Studies

Research involving marine mammals often requires costly field programs. This paper assessed whether the benefits of using cameras outweighs the implications of having personnel performing marine mammal detection in the field. The efficacy of video and still cameras to detect Indo-Pacific bottlenose dolphins (Tursiops aduncus) in the Fremantle Harbour (Western Australia) was evaluated, with consideration on how environmental conditions affect detectability. The cameras were set on a tower in the Fremantle Port channel and videos were perused at 1.75 times the normal speed. Images from the cameras were used to estimate position of dolphins at the water’s surface. Dolphin detections ranged from 5.6 m to 463.3 m for the video camera, and from 10.8 m to 347.8 m for the still camera. Detection range showed to be satisfactory when compared to distances at which dolphins would be detected by field observers. The relative effect of environmental conditions on detectability was considered by fitting a Generalised Estimation Equations (GEEs) model with Beaufort, level of glare and their interactions as predictors and a temporal auto-correlation structure. The best fit model indicated level of glare had an effect, with more intense periods of glare corresponding to lower occurrences of observed dolphins. However this effect was not large (-0.264) and the parameter estimate was associated with a large standard error (0.113). The limited field of view was the main restraint in that cameras can be only applied to detections of animals observed rather than counts of individuals. However, the use of cameras was effective for long term monitoring of occurrence of dolphins, outweighing the costs and reducing the health and safety risks to field personal. This study showed that cameras could be effectively implemented onshore for research such as studying changes in habitat use in response to development and construction activities.     

Homozygous M694V as a risk factor for amyloidosis in Turkish FMF patients

Secondary amyloidosis is the most severe complication of familial Mediterranean fever (FMF). Since the M694V mutation was associated with clinical severity, it was expected to be associated with amyloidosis as well. However, a number of contradicting reports have been published, especially pertinent to Turkish patients nearly 10 years ago. The aim of this study was to analyze recent data regarding the association between M694V mutation and amyloidosis among FMF patients in Turkey.We conducted a comprehensive review of the literature regarding the role of M694V mutation in the development of amyloidosis secondary to FMF. Twenty-seven papers from 20 centers including 3505 Turkish subjects were reviewed.Four-hundred patients had amyloidosis and homozygous M694V was detected in 189 (47%) of the 400 amyloidotic patients which was significantly higher than that in the FMF patients not developing amyloidosis (p < 0.0001).In the presented analysis we were able to reach a patient number of 400 which is much higher than all those published hitherto. Our findings confirmed that homozygous M694V is associated with amyloidosis in the Turkish population as well similar to Armenia, Israel, and Arabian countries. The necessity to treat asymptomatic or mildly symptomatic FMF patients with this genotype, even in countries where amyloidosis is rare, should be considered carefully.     

Seasonal Use of Latrines by Bobcats: Implications for Monitoring Programs

Latrines serve as important communication networks among felids for transmitting information relative to social dominance, reproductive status, and defense of hunting areas. During January 2011–August 2012, we monitored 10 bobcat (Lynx rufus) latrines in the northern Lower Peninsula (NLP) of Michigan, USA, using motion-sensitive cameras to estimate bobcat visitation and scat deposition rates among 3 biological seasons (mating, kitten-rearing, non-mating). Bobcat visitation rates differed among the 3 seasons. We found equal number of visits during the mating and kitten-rearing seasons, and lower visitation rates during the non-mating season. Scat deposition rates differed among the 3 seasons. We found a net gain of scats deposited during the mating and non-mating seasons, whereas there was a net loss of scats during the kitten-rearing season. An artificial latrine protocol we developed yielded visitation at 4 of 12 artificial latrine sites. Monitoring natural and artificial latrines during the mating and kitten-rearing seasons could provide valuable data for managing bobcat populations. © 2020 The Wildlife Society.     

Evidence that CT694 is a novel Chlamydia trachomatis T3S substrate capable of functioning during invasion or early cycle development

Chlamydia trachomatis is an obligate intracellular parasite, occupies a membrane-bound vacuole throughout development and is capable of manipulating the eukaryotic host by translocating effector molecules via a type III secretion system (T3SS). The infectious chlamydial elementary body (EB) is metabolically inactive yet possesses a functional T3S apparatus capable of translocating effector proteins into the host cell to facilitate invasion and other early cycle events. We present evidence here that the C. trachomatis protein CT694 represents an early cycle-associated effector protein. CT694 is secreted by the Yersinia T3SS and immunodetection studies of infected HeLa cultures indicate that CT694-specific signal accumulates directly adjacent to, but not completely overlapping with EBs during invasion. Yeast two-hybrid analyses revealed an interaction of CT694 with the repeat region and C-terminus of human AHNAK. Immunolocalization studies of CT694 ectopically expressed in HeLa cells were consistent with an interaction with endogenous AHNAK. Additionally, expression of CT694 in HeLa cells resulted in alterations in the detection of stress fibres that correlated with the ability of CT694 to interact with AHNAK. These data indicate that CT694 is a novel T3S-dependent substrate unique to C. trachomatis , and that its interaction with host proteins such as AHNAK may be important for aspects of invasion or development particular to this species.     

INTRODUCTION: Pheromones as Tools for Olfactory Research

We have long been fascinated by the unique ability of odorsto stir our emotions and to evoke long-forgotten memories, butcertain odors play a much more fundamental role in that theyvastly improve an organism's chances for reproductive successand survival. These odorants are called pheromones, a term commonlyapplied to semiochemicals that are released by one member ofa species and evoke a specific reaction or reactions from membersof the same species. Pheromones are known for both the specificityand the potency of their actions, which can be behavioral and/orneuroendocrinological. Pheromones can stimulate individualsto aggregate, to disperse, or to react defensively in the presenceof a predator, but they are probably best known for bringingthe sexes together. Some pheromones have also been found totrigger a dramatic release of pituitary hormones in severalvertebrate species. Although first identified in insects, morerecent studies show that sex pheromones influence the livesof a wide range of organisms, from microbes to man. The hormonally-derivedsex pheromones in teleost fish, and the airborne pheromonesof moths are two systems that illustrate how scientists haveused these specialized chemical signals as important tools toinvestigate the morphology, physiology and biochemistry of olfactory-receptorsystems, the mechanisms of odor-information processing in thebrain, and the diverse range of behaviors and endocrinologicalchanges associated with pheromonal communication. While ourfocus is on these two animal models, other examples, includingmammalian pheromone systems, are also discussed. Chem. Senses21: 241–243, 1996.     

Nicking Endonucleases as Unique Tools for Biotechnology and Gene Engineering

Russian Journal of Bioorganic Chemistry - Nicking endonucleases (NE) are a special group of the restriction endonucleases family. These unique enzymes catalyze the hydrolysis of only one DNA strand...     

Disability Measurement for Lymphatic Filariasis: A Review of Generic Tools Used within Morbidity Management Programs

Lymphatic filariasis (LF)-related disability affects 40 million people globally, making LF the leading cause of physical disability in the world. Despite this, there is limited research into how the impacts of LF-related disability are best measured. This article identifies the tools currently being used to measure LF-related disability and reviews their applicability against the known impacts of LF. The findings from the review show that the generic disability tools currently used by LF programs fail to measure the majority of known impacts of LF-related disability. The findings from the review support the development of an LF-specific disability measurement tool and raise doubt about the suitability of generic disability tools to assess disability related to neglected tropical diseases (NTDs) globally.     

Assessing Landbird Monitoring Programs and Demographic Causes of Population Trends

Abstract: Population trend data from the North American Breeding Bird Survey (BBS) have been used to identify conservation priorities and justify major conservation initiatives. Yet the BBS has been criticized for potential habitat bias and reliance on abundance indices to estimate trends. We compared 1992–2003 BBS trend estimates to trend estimates derived from bird-banding data collected as part of the Monitoring Avian Productivity and Survivorship (MAPS) program for 36 wood warbler species. Similarity in trends between the 2 monitoring programs at the survey-wide and program-wide scales suggested that each program can provide accurate trend information. The MAPS program, however, was designed primarily to complement (rather than duplicate) count-based efforts, such as the BBS, by providing estimates or indices of demographic rates. Demographic data from MAPS can be used to lend insight into proximate (demographic) causes of population trends and inform management. We illustrate this with analyses of 1992–2003 MAPS data for yellow warbler (Dendroica petechia). We used reverse-time capture-recapture models to evaluate importance of new recruits (including immigrating adults and young from the previous year) relative to surviving adults in explaining variation in trend among BBS physiographic strata. We included the number of young per adult captured (an index of productivity) as a covariate in models to assess effects of productivity on trends. Survival was the key demographic driver of recent population trends. Comparison of MAPS productivity indices and adult apparent survival rate estimates to BBS trend estimates largely confirmed this inference. We suggest that increased MAPS coverage, better coordination between MAPS and the BBS, and continued development of analytical methods that link the 2 programs will enhance the value of these monitoring efforts to land managers and conservation planners working at a variety of spatial scales.     

Strategies and Decision-Support Tools for Optimizing Long-Term Groundwater Monitoring Plans—MAROS 2.0

Existing long-term groundwater monitoring programs can be optimized to increase their effectiveness/efficiency with the potential to generate considerable cost savings. The optimization can be achieved through an overall evaluation of conditions of the contaminant plume and the monitoring network, focused spatial and temporal sampling analyses, and automated and efficient management of data, analyses, and reporting. Version 2.0 of the Monitoring and Remediation Optimization System (MAROS) software, by integrating long-term monitoring analysis strategies and innovative optimization methods with a data management, processing, and reporting system, allows site managers to quickly and readily develop cost-effective long-term groundwater monitoring plans. The MAROS optimization strategy consists of a hierarchical combination of analysis methods essential to the decision-making process. Analyses are performed in three phases: 1) evaluating site information and historical monitoring data to obtain local concentration trends and an overview of the plume status; 2) developing optimal sampling plans for future monitoring at the site with innovative optimization methods; and 3) assessing the statistical sufficiency of the sampling plans to provide insights into the future performance of the monitoring program. Two case studies are presented to demonstrate the usefulness of the developed techniques and the rigor of the software.     

人工转录因子研究进展

转录因子是真核表达调控中非常重要的一类反式作用因子,通常由DNA结合结构域与效应结构域两部分组成,研究发现这两个结构域可以各自独立发生作用。基于转录因子的这种结构特点,可以人为地选择针对特定序列的DNA结合结构域与具有特定作用的效应结构域构建人工转录因子。目前人工转录因子的DNA结合结构域多为C₂H₂ 型锌指结构,每一个锌指单元由大约30个氨基酸组成,识别DNA双螺旋大沟中相连的3bp序列,并可通过氢键作用与相应的碱基结合;多个锌指可以串联成簇,从而识别并结合较长的DNA序列区域。常见的人工转录因子的效应结构域有激活结构域以及抑制结构域,不同的效应结构域赋予人工转录因子不同的功能。目前人工转录因子已经在基础研究、药物设计以及基因治疗等领域得到了广泛的应用。     

P-type ATPases as drug targets: Tools for medicine and science

P-type ATPases catalyze the selective active transport of ions like H⁺, Na⁺, K⁺, Ca²⁺, Zn²⁺, and Cu²⁺ across diverse biological membrane systems. Many members of the P-type ATPase protein family, such as the Na⁺,K⁺-, H⁺,K⁺-, Ca²⁺-, and H⁺-ATPases, are involved in the development of pathophysiological conditions or provide critical function to pathogens. Therefore, they seem to be promising targets for future drugs and novel antifungal agents and herbicides. Here, we review the current knowledge about P-type ATPase inhibitors and their present use as tools in science, medicine, and biotechnology. Recent structural information on a variety of P-type ATPase family members signifies that all P-type ATPases can be expected to share a similar basic structure and a similar basic machinery of ion transport. The ion transport pathway crossing the membrane lipid bilayer is constructed of two access channels leading from either side of the membrane to the ion binding sites at a central cavity. The selective opening and closure of the access channels allows vectorial access/release of ions from the binding sites. Recent structural information along with new homology modeling of diverse P-type ATPases in complex with known ligands demonstrate that the most proficient way for the development of efficient and selective drugs is to target their ion transport pathway.     

Cloning of the Glyceraldehyde 3‐phosphate Dehydrogenase Gene of Flavescence dorée Phytoplasma and Development of Serological and Molecular Tools for Studying its Expression

The glyceraldehyde 3‐phosphate dehydrogenase (gapA) gene codes for a protein involved in the glycolytic pathway and is commonly used in Real‐Time RT‐PCR quantification studies as housekeeping gene. In this work we cloned and sequenced the full‐length gapA gene from Flavescence dorée phytoplasma (FDp). A ～35 kDa recombinant GapA protein was over‐expressed in Escherichia coli, purified and used as antigen to raise anti‐GapA rabbit polyclonal antibodies. The antiserum detected the GapA protein by western blot analysis of total protein extracts of FDp‐infected experimental host (Catharanthus roseus) and grapevine plants collected in the field. We also developed an FDp‐specific gapA Taqman Real‐Time RT‐PCR assay suitable for quantification overtime of gapA mRNA in infected plants.     

Prevalence and Risk Factors for Trachoma in Central and Southern Malawi

Background

Trachoma, one of the neglected tropical diseases is suspected to be endemic in Malawi. Objectives: To determine the prevalence of trachoma and associated risk factors in central and southern Malawi.

Methodology/Principal Findings

A population based survey conducted in randomly selected clusters in Chikwawa district (population 438,895), southern Malawi and Mchinji district (population 456,558), central Malawi. Children aged 1–9 years and adults aged 15 and above were assessed for clinical signs of trachoma. In total, 1010 households in Chikwawa and 1016 households in Mchinji districts were enumerated within 108 clusters (54 clusters in each district). A total of 6,792 persons were examined for ocular signs of trachoma. The prevalence of trachomatous inflammation, follicular (TF) among children aged 1–9 years was 13.6% (CI 11.6–15.6) in Chikwawa and 21.7% (CI 19.5–23.9) in Mchinji districts respectively. The prevalence of trachoma trichiasis (TT) in women and men aged 15 years and above was 0.6% (CI 0.2–0.9) in Chikwawa and 0.3% (CI 0.04–0.6) in Mchinji respectively. The presence of a dirty face was significantly associated with trachoma follicular (TF) in both Chikwawa and Mchinji districts (P<0.001).

Conclusion/Significance

Prevalence rates of trachoma follicles (TF) in Central and Southern Malawi exceeds the WHO guidelines for the intervention with mass antibiotic distribution (TF>10%), and warrants the trachoma SAFE control strategy to be undertaken in Chikwawa and Mchinji districts.