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The present study reviews Perinereis Group 2 species from the Eastern and South-eastern Asian seas based on morphological analysis of the types, non-types and original descriptions, and the use of molecular evidence (COI and 16S rDNA) from newly collected material. These species are characterized by having two bar-shaped paragnaths on pharyngeal area VI, which are often deemed conical when small and pointed, triggering misidentifications as to Neanthes species. New terminology and definition for this particular type of bars are proposed, and the generic position of some resembling Neanthes species is also re-assessed. Five species are transferred to Perinereis: Perinereis babuzai comb. nov., P. belawanensis comb. nov., P. kinmenensis comb. nov., P. shigungensis comb. nov. and P. vitabunda comb. nov. ‘Perinereis aibuhitensis’ species group is newly proposed by encompassing species having proximal dorsal ligule similar throughout the body, dorsal cirri short, and blades of heterogomph falcigers straight with long terminal tooth forming a distinct tendon. Perinereis belawanensis comb. nov., P. linea and P. vitabunda comb. nov. are redescribed. Perinereis linea is regarded as a senior synonym of Nereis (Neanthes) orientalis and Perinereis vancaurica tetradentata based on type material, whereas its exotic status in the Mediterranean Sea is questioned. An identification key to all currently valid species within Perinereis Group 2 is also provided.
The bone marrow (BM) is a frequent site of haematogenous spread for all types of cancer. Metastatic spread of disseminated tumour cells (DTCs) to the BM is detected in 0.2 to 12% of patients with solid tumours . The variability in incidence is related to the incidence of the primary tumour and its homing behaviour . Common primary tumours affecting the BM are listed below (Table 17.1).
Molecular diagnostics, i.e. the detection and analysis of disease-related changes of DNA or RNA, is becoming ever more important for the diagnosis of bone marrow (BM) neoplasms. In modern BM haematopathology, molecular diagnostics should always be part of an integrated diagnostic approach including clinical information, morphology and immunophenotyping. It is the responsibility of the haematopathologist to interpret the information gathered and to produce a final diagnosis. For this purpose, the practising haematopathologist must be familiar with the various molecular techniques needed and possess an in-depth knowledge of their applications for the diagnosis of BM neoplasms. The first part of this chapter focuses on the most important molecular techniques currently used in everyday diagnostics in the modern haematopathological laboratory. The second part highlights the major molecular and genetic aberrations of diagnostic value across the different haematological disease entities. Ideally the haematopathological laboratory should either be able to perform the relevant tests or be in close cooperation with a laboratory performing them for optimal diagnostics. Such cooperations also include multidisciplinary conferences, where clinicians and haematopathologists meet to discuss the diagnoses of afflicted patients.
Desoxyribosenucleic acid, DNA, and cellulose molecules self-assemble in aqueous systems. This aggregation is the basis of the important functions of these biological macromolecules. Both DNA and cellulose have significant polar and nonpolar parts and there is a delicate balance between hydrophilic and hydrophobic interactions. The hydrophilic interactions related to net charges have been thoroughly studied and are well understood. On the other hand, the detailed roles of hydrogen bonding and hydrophobic interactions have remained controversial. It is found that the contributions of hydrophobic interactions in driving important processes, like the double-helix formation of DNA and the aqueous dissolution of cellulose, are dominating whereas the net contribution from hydrogen bonding is small. In reviewing the roles of different interactions for DNA and cellulose it is useful to compare with the self-assembly features of surfactants, the simplest case of amphiphilic molecules. Pertinent information on the amphiphilic character of cellulose and DNA can be obtained from the association with surfactants, as well as on modifying the hydrophobic interactions by additives.
Chapter 3: This chapter charts the intersections between theatre and science and explores how experience and experiment are interlinked in each domain. Looking at three key scientific ideas and theatrical moments, the chapter draws out contextual aspects of the science that reflect the scientific concerns of their moments. Exploring first a play from the early years of the recent resurgence in the interest in theatre and science, the chapter investigates how the biological and medical sciences with their obvious link to genetic testing and human experience are represented, and moves on to consider how science, gender, and life become crystal clear in early twenty-first century theatre. It concludes by looking at how theatre is shaped by the experience of climate change and its science in the 2010s through two very different plays, one a staged lecture and the other a production whose deliberate excess results in an expansive ‘epic’ theatrical form that appears to take precedence over the science.
The aim of the present study was to investigate several common conditions that may potentially be correlated with follicular oxidative status during an intracytoplasmic sperm injection (ICSI) cycle and that include the serum oestrogen level on the day of oocyte pick-up, maternal age and pregnancy outcome. Patients that were enrolled in the study were classified randomly into three groups using their numerical order. The first group were classified based on maternal age (<35 and ≥35 years) (n = 398), the second group on the serum oestradiol (E2) level on the day of human chorionic gonadotropin (hCG) administration (levels >90th percentile and ≤ 90th percentile) (n = 491) and the third group on pregnancy outcome (positive/negative) (n = 376). The groups were matched for the other variables (stimulation protocol, dose of gonadotropin, duration of stimulation, antral follicle count, body mass index, basal follicle stimulating hormone (FSH), and E2 levels and day of hCG trigger) to prevent the possible contribution of those parameters to the results. Each group was matched for other variables (stimulation protocol, dose of gonadotrophin, duration of stimulation, antral follicle count, body mass index, basal FSH and E2 levels and day of hCG trigger) that may have affected the outcome, except for the parameter under investigation. Maternal age (P = 0.044,168 r = 0.418), oestrogen level on day of hCG administration (P = 0.001, r = 0.436) and pregnancy outcome (AUC = 0.65, P = 0.071) were found to be correlated with follicular oxidative status. The results obtained will help us to shield patients from possible situations that may cause oxidative stress and therefore adverse outcomes of an ICSI cycle.
There is a common misconception that our genomes - all unique, except for those in identical twins - have the upper hand in controlling our destiny. The latest genetic discoveries, however, do not support that view. Although genetic variation does influence differences in various human behaviours to a greater or lesser degree, most of the time this does not undermine our genuine free will. Genetic determinism comes into play only in various medical conditions, notably some psychiatric syndromes. Denis Alexander here demonstrates that we are not slaves to our genes. He shows how a predisposition to behave in certain ways is influenced at a molecular level by particular genes. Yet a far greater influence on our behaviours is our world-views that lie beyond science - and that have an impact on how we think the latest genetic discoveries should, or should not, be applied. Written in an engaging style, Alexander's book offers tools for understanding and assessing the latest genetic discoveries critically.
In the chapter “The Biotechnology Sector – Therapeutics”, the author covers a wide range of topics summarizing the significant role that the formation and growth of the biotechnology sector has played in the entire biopharmaceutical industry. The chapter begins with a bit of history, from the earliest days of how genetic engineering gave birth to this sector, and takes the reader through an overview of biotechnology as it exists today and how the growing innovation in science over the years has been able to both drive the sector and have a tremendous impact on healthcare overall. There is a particular focus on describing various types of innovation which have played a huge role in driving product development in the broader biopharmaceutical industry. Later in the chapter, there is a focus on many of the business aspects of the sector, as drug development in biotechnology requires enormous amounts of capital for success. The author outlines many of the key issues related to different business and financing models that we see across the sector, in addition to the unique management issues in small biotechnology companies. There is significant description and explanation of the symbiotic relationship between the larger pharmaceutical companies and smaller biotechnology start-ups with a focus on how they help each other to bring transformative medicines to patients. The chapter concludes with a discussion about international and regulatory aspects impacting the sector. Overall the author tells the story of the birth and growth of this exciting sector, and its impact on patients and drug development over the last forty years, well substantiated with current data to build the case for how biotechnology today plays a major role in driving one of the most important and exciting technological industries of our time.
Genetic studies provide a compelling story of gene influences on intelligence, and neuroimaging studies provide insights about relevant brain structure and function. Polygenetic scores based on DNA and brain connectivity patterns based on neuroimaging are beginning to show correlations with individual differences in intelligence. Imaging studies also provide insights on specific brain networks related to intelligence, especially the PFIT model. The concept of brain efficiency is now being explored at the network and the dendrite levels. As we push inexorably deeper into the brain from cortex to neurons to synapses, we are at the threshold of developing a molecular biology of intelligence based both on gene expression related to brain development and function, and on the cascades of neurobiological events at the neuron and synapse levels. As prediction advances and the biological mechanisms underlying intelligence are identified, a major step will be manipulation of those mechanisms to enhance intelligence. That is why the study of intelligence has never been more exciting or important.
Species of the allocreadiid genus Creptotrema are parasites of freshwater fishes in the Americas. Species in the genus possess one pair of muscular oral lobes on the oral sucker. Currently, the genus contains eight species, six distributed in South America, one in Middle America and one in North America. Genetic data are only available for the North American species, Creptotrema funduli, a parasite of fundulids originally described from Oneida Lake, New York State. In this study, we obtained 28S ribosomal DNA sequences of trematodes morphologically similar to Creptotrema agonostomi from the mountain mullet, Dajaus monticola, across a wide geographical range in Middle America. Our molecular phylogenetic analyses showed that (1) the genus Creptotrema, as currently conceived, is not monophyletic; (2) the allocreadiids in mountain mullets should be re-allocated in the genus Pseudoparacreptotrema; and (3) the allocreadiid trematodes from D. monticola across Middle America represent four morphologically similar species, three of which can be distinguished genetically. These three new species are described herein using an integrative taxonomy approach. We contend that accurate estimates of species diversity and phylogenetic relationships among allocreadiids, and most likely other species of trematodes, necessarily require an integrative taxonomy approach that should consider at least DNA sequences and scanning electron microscopy.
Biobanks are a valuable resource for creating advancements in science through cutting-edge omics research. Twin research methods allow us to understand the degree to which genetics and environmental factors contribute to health outcomes.
The Sri Lankan Twin Registry biobank (SLTR-b) was established in 2015 as part of Colombo Twin and Singleton Follow-up Study. Venous blood and urine were collected from twins and comparative sample of singletons for clinical investigations and biobanking.
The SLTR-b currently houses 3369 DNA and serum samples. Biobank specimens are linked to longitudinal questionnaire data, clinical investigations, anthropometric measurements, and other data.
The SLTR-b aims to address gaps in health and genetics research. It will provide opportunities for academic collaborations, local and international, and capacity building of future research leaders in twin and omics research. This paper provides a cohort profile of the SLTR-b and its linked data, and an overview of the strategies used for biobanking.
Neospora caninum is a commonly diagnosed cause of reproductive losses in farmed ruminants worldwide. This study examined 495 and 308 samples (brain, heart and placenta) which were collected from 455 and 119 aborted cattle and sheep fetuses, respectively. DNA was extracted and a nested Neospora ITS1 PCR was performed on all samples. The results showed that for bovine fetuses 79/449 brain [17.6% (14.2–21.4)], 7/25 heart [28.0% (12.1–49.4)] and 5/21 placenta [23.8% (8.2–47.2)] were PCR positive for the presence of Neospora DNA. Overall 82/455 [18.0% (14.6–21.7)] of the bovine fetuses tested positive for the presence of N. caninum DNA in at least one sample. None (0/308) of the ovine fetal samples tested positive for the presence of Neospora DNA in any of the tissues tested. The results show that N. caninum was associated with fetal losses in cattle (distributed across South-West Scotland), compared to sheep in the same geographical areas where no parasite DNA was found. Neospora is well distributed amongst cattle in South-West Scotland and is the potential cause of serious economic losses to the Scottish cattle farming community; however, it does not appear to be a problem amongst the Scottish sheep flocks.
While some single-celled intestinal parasites are direct causes of diarrhoea and other types of intestinal pathology, the impact of other gut micro-eukaryotes on human health remains elusive. The fact that some common luminal intestinal parasitic protists (CLIPPs) have lately been found more often in healthy than in diseased individuals has fuelled the hypothesis that some parasites might in fact be protective against disease. To this end, the use of new DNA technologies has helped us investigate trans-kingdom relationships in the gut. However, research into these relationships is currently hampered by the limited data available on the genetic diversity within the CLIPPs genera, which results in limited efficacy of publicly available DNA sequence databases for taxonomic annotation of sequences belonging to the eukaryotic component of the gut microbiota. In this paper, I give a brief overview of the status on CLIPPs in human health and disease and challenges related to the mapping of intestinal eukaryotic diversity of the human gut.
Despite the reduction in the prevalence of soil-transmitted helminthiases in many regions of the world, morbidity rates remain high in some rural regions. The Kato–Katz technique is a simple, inexpensive and field-applicable tool commonly used for the diagnosis and worm-burden characterization of these infections. Molecular studies have revolutionized our understanding of the epidemiology and evolutionary genetics of parasites. In this study we recovered helminthic DNA from Kato–Katz slides (n = 93) prepared in 2011 in the Brazilian Amazon. We achieved DNA recovery by polymerase chain reaction (PCR) in 84% of cases for Ascaris sp. and 75% of cases for hookworms. The sequencing confirmed the specific species of the amplicons. The slides stored for a few years could be analysed using this methodology, allowing access to DNA from a large collection of samples. We must consider the Kato–Katz thick smears as a source of helminth DNA. This can significantly reduce logistical difficulties in the field in terms of obtaining, preserving, transporting and initial processing of samples.
With the push towards control and elimination of soil-transmitted helminthiasis and schistosomiasis in low- and middle-income countries, there is a need to develop alternative diagnostic assays that complement the current in-country resources, preferably at a lower cost. Here, we describe a novel high-resolution melt (HRM) curve assay with six PCR primer pairs, designed to sub-regions of the nuclear ribosomal locus. Used within a single reaction and dye detection channel, they are able to discriminate Ancylostoma duodenale, Necator americanus, Strongyloides stercoralis, Ascaris lumbricoides, Trichuris trichiuria and Schistosoma spp. by HRM curve analysis. Here we describe the primers and the results of a pilot assessment whereby the HRM assay was tested against a selection of archived fecal samples from Ghanaian children as characterized by Kato–Katz and real-time PCR analysis with species-specific TaqMan hydrolysis probes. The resulting sensitivity and specificity of the HRM was 80 and 98.6% respectively. We judge the assay to be appropriate in modestly equipped and resourced laboratories. This method provides a potentially cheaper alternative to the TaqMan method for laboratories in lower resource settings. However, the assay requires a more extensive assessment as the samples used were not representative of all target organisms.
In this study, nano-hydroxyapatite (n-HAp) of average crystallite size ∼8.15 ± 4 nm of hexagonal geometry with size ranging between 14 and 50 nm was synthesized in laboratory at room temperature by using suitable sources of calcium and phosphate ions and using triethanolamine. Mesoporous bioactive glass (MBG) was synthesized by using cationic surfactant cetyl trimethyl ammonium bromide of the SiO2–CaO–P2O5 glass system. After calcination at 650 °C, MBG powders were having a zeta potential of −16.5 mV (pH ∼9.1), median particle size ∼75 nm, and specific surface area 473.2 m2/g. An aqueous suspension of DNA was used to disperse both n-HAp and MBG and further subjected for analysis including absorbance, circular dichroism spectroscopy, UV-melting, and isothermal titration calorimetry. Absorbance spectroscopy indicated that an equilibrium binding was obtained between both materials and DNA in solution phase. Due to the addition of the nanomaterial, molar ellipticity of DNA was changed revealing that the materials were interacted with DNA. From UV melting characterization, there is a shifting of the melting temperature of DNA in the presence of MBG and n-HAp, respectively, suggesting that the nanoparticles stabilized DNA helix to a considerable extent.
Molecular methods have been developed for the detection and quantification of Trypanosoma cruzi DNA in blood samples from patients with Chagas disease. However, aspects of sample processing necessary for quantitative real-time PCR (qPCR), such as the addition of guanidine hydrochloride to whole blood samples, may limit timely access to molecular diagnosis. We analysed 169 samples from serum and guanidine-EDTA blood (GEB) obtained from patients in acute and chronic phases of Chagas disease. We applied qPCR targeted to the satellite DNA region. Finally, we compared the parasite loads and cycle of threshold values of the qPCR. The results confirmed the usefulness of serum samples for the detection and quantification of parasite DNA in patients with Chagas disease, especially in the acute phase. However, the parasite loads detected in serum samples from patients in the chronic phase were lower than those detected in GEB samples. The epidemiological implications of the findings are herein discussed.
Toxoplasma gondii infections are acquired through the ingestion of oocysts present in the environment. However, there is no data about their occurrence in the air or about airborne transmission of these infections. In the present paper, we report on the identification of T. gondii using rapid molecular detection methods, supported by microscopic analysis, in environmental air samples. A total of 71 samples were collected, using gelatine filters, from kitchen gardens, recreational areas and sandpits located in northern and north-eastern Poland. Material recovered from the filters was analysed using real-time PCR and loop-mediated isothermal assays targeting the T. gondii B1 gene. Toxoplasma gondii DNA was found in two samples, as confirmed by both molecular assays. Genotyping at the SAG2 locus showed Toxoplasma SAG2 type I. Moreover, the presence of T. gondii oocysts was confirmed in one of the positive samples with the use of microscopy. The results showed that T. gondii may be present in environmental air samples and that respiratory tract infections may play a role in the high prevalence of toxoplasmosis in humans and animals. To the best of our knowledge, this is the first epidemiological evidence that oro-fecal and foodborne toxoplasmosis may be traceable to an airborne respiratory origin and that this may represent a new, previously unknown transmission route for this disease.
Babesia are intraerythrocytic parasites of importance worldwide within the fields of human and veterinary medicine, as some Babesia sp., including Babesia microti are potentially zoonotic and can cause fatal disease in both humans and animals. The aims of this study were to use a nested PCR (amplifying the 18S rRNA gene) to determine the presence and species of Babesia parasite DNA found in blood (n = 47) and spleen (n = 47) samples collected from Eurasian badgers (Meles meles) in Scotland. The results showed 28/47 (59·6%) blood and 14/47 (29·8%) spleen samples tested positive for the presence of Babesia DNA. Initial sequence analysis of the Babesia DNA identified three distinct sequence types (submitted to GenBank KX528553, KX528554 and KX528555), which demonstrated ⩾99% identity to Babesia sp. parasites previously identified in badgers in Spain (KT223484 and KT223485). Phylogenetic analysis showed that the three isolates are closely related to Babesia annae, B. microti and other Piroplasmida species found in wildlife. Further sequence analysis of the samples demonstrated that the badgers were routinely infected with more than one parasite isolate and there was also evidence of genetic recombination between the Babesia parasite isolates (submitted to GenBank KY250472 – KY250477)
Previous investigations suggested that elevated cell-free DNA (cfDNA) can indicate non-healthy states. However, the potential association between cfDNA seminal plasma levels and fertility sperm parameters has not yet been determined. Therefore, the present study evaluated the association between seminal cfDNA levels and sperm fertility criteria to determine the use of seminal cfDNA quantification. An in vivo protocol quantified cfDNA levels of semen samples obtained from 163 male patients using fluorescent PicoGreen dye staining. To confirm if semen cfDNA quantification is realistic, an in vitro complementary test was performed using three or four semen samples. The fresh sperm samples were exposed to paraquat that generates high levels of superoxide anion causing oxidative stress and cell mortality. The results showed significant association between dsDNA levels and several sperm fertility parameters, such as low viability and alterations of motility and morphology. The in vitro analysis confirmed the association between dsDNA levels and sperm viability. Together, these results suggest that dsDNA levels could be an important biomarker to test sperm fertility.