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\"Timely and beautifully written, New England Beyond Criticism provides a passionate defense of the importance of the literature of New England to the American literary canon, and its impact on the development of spirituality, community, and culture in America. An exploration and defense of the prominence of New England's literary tradition within the canon of American literature. Traces the impact of the literature of New England on the development of spirituality, community, and culture in America. Includes in-depth studies of work from authors and poets such as William Bradford, Emily Dickinson, Robert Frost, Henry David Thoreau, Susan Howe, and Marilynne Robinson. Examines the place and impression of New England literature in the nation's intellectual history and the lives of its readers\"-- Provided by publisher.

The diagnostics of ruminant parasites remains one of the cornerstones for parasite control best practices. Field veterinarians have several techniques at their disposal (fecal egg count, coproculture, FAMACHA®, plasma pepsinogen, ELISA- Ostertagia , ELISA- Fasciola , Baermann and ELISA-Lungworm) for the identification and/or quantification of gastrointestinal nematodes, lungworms and liver fluke infecting small ruminants and cattle. Each of these diagnostic tools has its own strengths and weaknesses and is more appropriate for a specific production operation and/or age of the animal (young and adults). This review focuses on the usability and interpretation of the results of these diagnostic tools. The most advanced technical information on sampling, storage, advantages and limitations of each tool for different types of production operations and animal categories is provided. Graphical abstract

Rapid and accurate detection of fentanyl in unknown products and aqueous samples can find the unexpected existence of the toxic chemical and assist law enforcement. In this paper, novel colorimetric biosensors for fentanyl and one of its metabolites, norfentanyl, are designed and fabricated by incorporating an enzyme‐linked immunosorbent assay (ELISA) onto three dimensionally macroporous framework melamine foam (MF). The MF‐based ELISA sensors (f‐ELISA) have performance features of ultrahigh sensitivity, selectivity, and on‐site detection without using any laboratory instruments. The lowest visually distinguishable concentration of fentanyl in wastewater is 0.005 mg L−1 by naked eye and can be further improved to 0.001 mg L−1 with the aid of a smartphone. Furthermore, by increasing sample volume, the sensitivity of the f‐ELISA sensor can reach naked‐eye detection of fentanyl at concentrations as low as 0.0005 mg L−1, a volume‐responsive signal enhancement, and a special structural advantage provided by the macroporous structure of the foam. The norfentanyl f‐ELISA sensor can reveal a lowest visually distinguishable concentration of 0.001 mg L−1 in PBS buffer for both naked‐eye and smartphone analysis. The new biosensors can overcome limitations of current techniques in detecting fentanyl and its derivatives in fluids and are suitable for on‐site use. A foam‐based ELISA (f‐ELISA) platform was developed using chemically modified melamine foam to enable rapid, ultrasensitive, and selective colorimetric detection of fentanyl and norfentanyl.

ABSTRACT Background Ethiopia has a significant number of sheep and goats, though the benefit gained is little because of several diseases, including Peste des Petits Ruminants (PPR). PPR is a highly contagious and economically important transboundary disease of small ruminants, associated with high morbidity and mortality rates. Objectives A cross‐sectional study was conducted to estimate seroprevalence of PPR and identify associated risk factors in four Kebeles of the Pawe district, Northwest Ethiopia. Methods A total of 334 serum samples (146 from goats and 188 from sheep) were collected and tested for PPRV antibodies using competitive ELISA (c‐ELISA). Results Out of a total of 334 blood sera from sheep and goats tested for PPR virus antibodies, 236 (70.7%) were positive. There was no significant difference in prevalence between sheep (71.3%) and goats (69.9%). A significantly higher PPRV seroprevalence was observed in female sheep (75.7%) compared to male sheep (55%). Similarly, female goats had a higher prevalence (74.4%) than male goats (48%) (p value = 0.01). The prevalence was also significantly higher in lactating female sheep (81.8%) and goats (83.3%). Age‐wise, seroprevalence was higher in older sheep (82.2%, odds ratios [OR] = 4.4) and older goats (84.1%, OR = 5.4) than their younger counterparts. The prevalence was higher in Almu Kebele (82.5%) (χ2 = 7.5, p value = 0.05). Flock size, age, body condition, origin and the introduction of new animals were identified as potential risk factors associated with the prevalence. Conclusions PPR is highly prevalent in the district, indicating significant circulation of the virus in the area. This high prevalence, combined with the regular movement of animals and shared grazing lands in the area, poses a risk for the further spread of the disease. Therefore, priority should be given to the area in control and eradication efforts, along with restrictions on animal movement and targeted vaccination. GRAPHICAL ABSTRACT ❖ The overall antibody seroprevalence of PPRV in the district was 70.7% (236/334) with nonsignificant difference in the prevalence between sheep (71.3%) and goats (69.9%). ❖ Highest Kebele‐wise overall antibody seroprevalence of PPRV was recorded in Almu (82.5%), followed by Mender 14 (68.7%), and the lowest seroprevalence was recorded from Mender 23/45 Kebele (64.6%). ❖ Age, physiological status, sex, Kebele and housing system of the shoats were significantly associated with seropositivity of small ruminants to PPRV antibodies.

Programmed cell death is considered a key player in a variety of cellular processes that helps to regulate tissue growth, embryogenesis, cell turnover, immune response, and other biological processes. Among different types of cell death, apoptosis has been studied widely, especially in the field of cancer research to understand and analyse cellular mechanisms, and signaling pathways that control cell cycle arrest. Hallmarks of different types of cell death have been identified by following the patterns and events through microscopy. Identified biomarkers have also supported drug development to induce cell death in cancerous cells. There are various serological and microscopic techniques with advantages and limitations, that are available and are being utilized to detect and study the mechanism of cell death. The complexity of the mechanism and difficulties in distinguishing among different types of programmed cell death make it challenging to carry out the interventions and delay its progression. In this review, mechanisms of different forms of programmed cell death along with their conventional and unconventional methods of detection of have been critically reviewed systematically and categorized on the basis of morphological hallmarks and biomarkers to understand the principle, mechanism, application, advantages and disadvantages of each method. Furthermore, a very comprehensive comparative analysis has been drawn to highlight the most efficient and effective methods of detection of programmed cell death, helping researchers to make a reliable and prudent selection among the available methods of cell death assay. Conclusively, how programmed cell death detection methods can be improved and can provide information about distinctive stages of cell death detection have been discussed.

During biologics development, manufacturers must demonstrate clearance of host cell impurities and contaminants to ensure drug purity, manufacturing process consistency, and patient safety. Host cell proteins (HCPs) are a major class of process-related impurities and require monitoring and documentation of their presence through development and manufacturing. Even in residual amounts, they are known to affect product quality and efficacy as well as patient safety. HCP analysis using enzyme-linked immunosorbent assay (HCP-ELISA) is the standard technique, due to its simple handling, short analysis time, and high sensitivity for protein impurities. Liquid chromatography mass spectrometry (LC–MS) is an orthogonal method for HCP analysis and is increasingly included in regulatory documentation. LC–MS offers advantages where HCP-ELISA has drawbacks, e.g., the ability to identify and quantify individual HCPs. This article summarizes the available knowledge about monitoring HCPs in biologics and presents the newest trends in HCP analysis with current state-of-the-art HCP measurement tools. Through case studies, we present examples of HCP control strategies that have been used in regulatory license applications, using an MS-based coverage analysis and HCP-ELISA and LC–MS for HCP quantification. This provides novel insight into the rapid evolving strategy of HCP analysis. Improvements in technologies to evaluate HCP-ELISA suitability and the implementation of orthogonal LC–MS methods for HCP analysis are important to rationally manipulate, engineer, and select suitable cell lines and downstream processing steps to limit problematic HCPs.

Coccidioidomycosis is often diagnosed with a collection of tests that rely on the patient's ability to mount an immune response to the fungus (antibody-based diagnostics), making diagnosis of this infection challenging. Here we present an antigen-based assay that detects and quantifies coccidioidal chitinase-1 (CTS1) in human serum.BACKGROUNDCoccidioidomycosis is often diagnosed with a collection of tests that rely on the patient's ability to mount an immune response to the fungus (antibody-based diagnostics), making diagnosis of this infection challenging. Here we present an antigen-based assay that detects and quantifies coccidioidal chitinase-1 (CTS1) in human serum.An inhibition-based enzyme-linked immunoassay (ELISA) was developed that utilizes a monoclonal antibody specific for coccidioidal CTS1. CTS1 was quantified in commercial antigen preparations using recombinant CTS1 as a standard. Sera from 192 individuals from an endemic area were tested, which included 78 patients (40.6%) with proven or probable coccidioidomycosis.METHODSAn inhibition-based enzyme-linked immunoassay (ELISA) was developed that utilizes a monoclonal antibody specific for coccidioidal CTS1. CTS1 was quantified in commercial antigen preparations using recombinant CTS1 as a standard. Sera from 192 individuals from an endemic area were tested, which included 78 patients (40.6%) with proven or probable coccidioidomycosis.The quantity of CTS1 in diagnostic commercial antigen preparations from different suppliers varied. CTS1 antigenemia was detected in 87.2% of patients with proven or probable coccidioidomycosis. Specificity was determined to be 96.94% using serum from individuals who reside in the Phoenix, Arizona area who did not have coccidioidomycosis. Levels of CTS1 correlated with low- and high-titer serology from patients with a coccidioidomycosis diagnosis.RESULTSThe quantity of CTS1 in diagnostic commercial antigen preparations from different suppliers varied. CTS1 antigenemia was detected in 87.2% of patients with proven or probable coccidioidomycosis. Specificity was determined to be 96.94% using serum from individuals who reside in the Phoenix, Arizona area who did not have coccidioidomycosis. Levels of CTS1 correlated with low- and high-titer serology from patients with a coccidioidomycosis diagnosis.Since the CTS1 inhibition ELISA described in this report does not depend on the host immune response, it is a promising diagnostic tool to aid in diagnosis and disease monitoring of coccidioidomycosis.CONCLUSIONSSince the CTS1 inhibition ELISA described in this report does not depend on the host immune response, it is a promising diagnostic tool to aid in diagnosis and disease monitoring of coccidioidomycosis.

The safety of gluten-free foods is essential for celiac disease (CD) patients to prevent serious complications. Enzyme-linked immunosorbent assays (ELISAs) are recommended for gluten analysis to monitor the compliance of gluten-free products to the Codex threshold of 20 mg gluten/kg. However, due to the specific features of each gluten ELISA test kit, the results often deviate systematically and largely depend on the characteristics of the antibody. This comprehensive study assessed the specificities and sensitivities of three monoclonal (R5, G12, and Skerritt) and two polyclonal antibodies to the alcohol-soluble prolamin and alcohol-insoluble glutelin fractions of gluten from wheat, rye, and barley, all of which harbor CD-active epitopes. Reversed-phase high-performance liquid chromatography served as independent reference method to quantify gluten protein concentrations and allow comparisons of different gluten fractions within one kit and between kits. Wheat prolamins were detected quite accurately by all antibodies, but high variability between antibody specificities and sensitivities was observed for rye and barley prolamins and rye glutelins, and the largest discrepancies were found for wheat and barley glutelins. The gluten content (sum of prolamins and glutelins) was either overestimated up to six times (rye) or underestimated up to seven times (barley). Overestimation of gluten contents may unnecessarily limit the availability of gluten-free products, but underestimation represents a serious health risk for CD patients. It is important to consider these differences between antibodies used in kits and consider what each kit is capable of measuring, especially with samples where the source of gluten is unknown.

Neurocysticercosis prevalence estimates often are based on serosurveys. However, assessments of Taenia solium seropositivity durability in patients with various neurocysticercosis types are lacking. We optimized a triplex serologic ELISA by using synthetic GP50, T24H, and Ts18var3 antigens for T. solium. We used that assay to test sequential serologic responses over several years after neurocysticercosis cure in 46 patients, 9 each with parenchymal or ventricular neurocysticercosis and 28 with subarachnoid disease. Triplex results were concordant with 98% of positive and 100% of negative enzyme-linked immunoelectrotransfer blots. Eight years after neurocysticercosis cure, 11.1% of patients with parenchymal, 47.3% with subarachnoid, and 41.7% with ventricular disease were still seropositive. Median time to seroreversion after cure in this cohort in a T. solium nonendemic area was 2 years for parenchymal disease, 4 years for ventricular disease, and 8 years for subarachnoid disease. Our findings can inform epidemiologic models that rely on serosurveys to estimate disease burden.

Background Leishmaniosis caused by Leishmania infantum , L. major and L. tropica is endemic in Morocco. Growing evidence of both human and canine Leishmania infections in urban centres has been reported. Since many forms of the disease are zoonotic, veterinarians play an important role in leishmaniosis control by intervening at the parasite host level. This study aimed to bring together One Health principles to connect canine and feline leishmaniosis epidemiology within urban centres of Morocco (Rabat and Fez) and assess the level of awareness of Moroccan veterinarians about facing this threat. Methods A molecular survey was conducted for Leishmania DNA detection in canine ( n =  155) and feline ( n  = 32) whole-blood samples. Three conventional polymerase chain reaction (PCR) protocols were implemented. The first PCR aimed at identifying infected animals by targeting Leishmania spp. kinetoplast minicircle DNA (kDNA). The second and third PCR targeted the Leishmania internal transcribed spacer region (ITS-1) and the Leishmania small subunit ribosomal RNA (SSUrRNA) gene, respectively, aiming at identification of the infecting species after Sanger sequencing-positive amplicons. Total immunoglobulin G (IgG) against Leishmania spp. was evaluated in 125 dogs by enzyme-linked immunosorbent assays (ELISA) using an in-house protocol, including three Leishmania -specific antigens (SPLA, rKDDR and LicTXNPx). Sera from 25 cats were screened for total IgG to Leishmania spp. by an indirect immunofluorescence antibody test (IFAT). An online questionnaire was presented to Moroccan veterinarians addressing their knowledge and practices towards animal leishmaniosis. Results Overall, 19.4% of the dogs tested positive for Leishmania kDNA and ITS-1 and sequencing revealed infection with L. infantum among PCR-positive dogs. These animals presented a wide range of ELISA seropositivity results (16.7%, 34.9% and 51.6%) according to the tested antigens (rKDDR, SPLA and LicTXNPx, respectively). Use of kDNA-PCR revealed 12.5% cats positive to Leishmania spp. otherwise found to be seronegative by IFAT. Conclusions A considerable prevalence of infection was identified in dogs from urban centres of Morocco. Additionally, this is the first report of feline infection with Leishmania spp. in this country and in urban settings. Moroccan veterinarians are aware that animal leishmaniosis is endemic in Morocco, representing a public health threat, and are knowledgeable about canine leishmaniosis diagnosis and treatment. Graphical Abstract