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1.
Regul Toxicol Pharmacol ; 147: 105571, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38244664

ABSTRACT

The World Health Organization (WHO) assesses potential health risks of dioxin-like compounds using Toxic Equivalency Factors (TEFs). This study systematically updated the relative potency (REP) database underlying the 2005 WHO TEFs and applied advanced methods for quantitative integration of study quality and dose-response. Data obtained from fifty-one publications more than doubled the size of the previous REP database (∼1300 datasets). REP quality and relevance for these data was assessed via application of a consensus-based weighting framework. Using Bayesian dose-response modeling, available data were modeled to produce standardized dose/concentration-response Hill curves. Study quality and REP data were synthesized via Bayesian meta-analysis to integrate dose/concentration-response data, author-calculated REPs and benchmark ratios. The output is a prediction of the most likely relationship between each congener and its reference as model-predicted TEF uncertainty distributions, or the 'best estimate TEF' (BE-TEF). The resulting weighted BE-TEFs were similar to the 2005 TEFs, though provide more information to inform selection of TEF values as well as to provide risk assessors and managers with information needed to quantitatively characterize uncertainty around TEF values. Collectively, these efforts produce an updated REP database and an objective, reproducible approach to support development of TEF values based on all available data.


Subject(s)
Dioxins , Polychlorinated Biphenyls , Animals , Dioxins/toxicity , Bayes Theorem , Mammals
2.
Clin Radiol ; 78(12): e941-e949, 2023 12.
Article in English | MEDLINE | ID: mdl-37788968

ABSTRACT

AIM: To investigate how magnetic resonance imaging (MRI) examinations are protocolled in tertiary paediatric neuroradiology centres around the UK for some of the more common presentations encountered in paediatric neuroradiology, and to identify any variations of note. MATERIALS AND METHODS: All 19 UK tertiary paediatric neuroradiology centres registered with the British Society of Neuroradiologists-Paediatric Group were contacted and asked if they could provide a copy of their standard MRI protocols. Twelve responded (63%) and 10 of the more common presentations were selected and the standard acquired sequences obtained at each participating centre were compared. Where available the collated protocols were also compared against current published guidance. RESULTS: The basic sequences carried out by centres around the UK are similar; however, there are lots of variations overall. The only standardised protocol currently being implemented nationally in paediatric imaging is that for brain tumours. Otherwise, chosen protocols are generally dependent on the preferences and technical capabilities of individual centres. Suggested published protocols also exist for non-accidental injury (NAI), multiple sclerosis, epilepsy, and head and neck imaging. CONCLUSIONS: The differences in MRI protocolling depend in part on technical capabilities and in part on the experience and preferences of the paediatric neuroradiologists at each centre. For most presentations, there is no consensus as to what constitutes the perfect protocol. The present results will be useful for specialist centres who may wish to review their current protocols, and for more generalist centres to use as a reference to guide their MRI protocolling.


Subject(s)
Brain Neoplasms , Hospitals, Pediatric , Child , Humans , Brain Neoplasms/diagnostic imaging , Magnetic Resonance Imaging/methods , Tertiary Care Centers , United Kingdom
3.
Osteoporos Int ; 33(6): 1347-1355, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35088102

ABSTRACT

In this retrospective cohort study, men and women with eating disorders (n = 8867) had higher risk of injurious falls and hip fractures than age, sex, and county-matched controls (n = 88670). INTRODUCTION: Eating disorders have been associated with decreased bone mineral density and increased fracture risk, but the association with fall injuries without fracture has not previously been investigated. Furthermore, fracture risk in men with eating disorders has been insufficiently studied. METHODS: In the present study, 8867 patients (9.4% men) with a diagnosed eating disorders and 88670 age-, sex-, and county-matched controls were investigated. RESULTS: The mean (standard deviation) age of the patients and controls was 41.6 (13.7) years and the follow-up time 9.6 (5.2, 14.4) years (median, interquartile range) for patients and 10.1 (5.5, 14.2) years for controls. The proportions of injurious falls without fracture (17.3% vs. 9.0%) and of hip fracture (1.6% vs. 0.7%) were substantially greater in patients with an eating disorder than in their corresponding population controls. In an unadjusted Cox proportional hazards model, individuals with an eating disorder had a higher risk of injurious falls without fracture (Hazard ratio (HR) 95% confidence interval (CI): 2.07 (1.96-2.18), and hip fracture (HR 2.30 (1.92-2.75)) than the risk observed in the controls. The HR for any investigated outcome associated with an eating disorder did not differ by sex or age (interaction term p > 0.10). The risk of injurious falls without fracture and hip fracture was increased in both women (HR 2.07 (1.95-2.19) and HR 2.41 (1.98-2.93), respectively) and men (HR 2.09 (1.76-2.49) and HR 1.84(1.12-3.02), respectively), with an eating disorder. CONCLUSION: The risk of injurious falls without fracture and of hip fracture is increased in both women and men with eating disorders, indicating measures to prevent both falls and fractures are important in these patients, regardless of age and sex.


Subject(s)
Feeding and Eating Disorders , Hip Fractures , Bone Density , Child , Feeding and Eating Disorders/complications , Feeding and Eating Disorders/epidemiology , Female , Hip Fractures/epidemiology , Hip Fractures/etiology , Humans , Male , Retrospective Studies , Risk Factors , Sweden/epidemiology
4.
Clin Exp Dermatol ; 47(1): 37-42, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34160837

ABSTRACT

BACKGROUND: Skin conditions have a large emotional, psychological and psychiatric impact on the individual. The All-Party Parliamentary Group on Skin (APPGS) commissioned a qualitative survey to further explore this relationship alongside the experiences of those accessing services in relation to these difficulties in the UK. AIM: To examine the experiences of UK individuals living with a skin condition, and their views of seeking and receiving psychological treatment. This survey formed part of the evidence collected in the preparation of the APPGS Mental Health and Skin Disease report. METHODS: A free-text electronic survey was widely distributed by professional bodies and skin-related charities. Responses were analysed using descriptive thematic analysis and descriptive statistics. Data for each question were classified and labelled, leading to the development of a coding frame. Inter-rater reliability was assessed using Cohen kappa statistic. RESULTS: In total, 544 participants (84% female) completed the survey. The majority of respondents had inflammatory skin diseases such as eczema (43%) or psoriasis (33%). The thematic analysis revealed five key themes associated with impact on mood; impact of intimacy; impact on activities of daily living; lack of recognition from others of the impact; and lack of accessible services. CONCLUSION: The survey demonstrates that there is an urgent need to improve both awareness of the impact that skin conditions can have, and for the provision of psychological services to address this impact.


Subject(s)
Health Services Accessibility , Psychosocial Support Systems , Skin Diseases/psychology , Adult , Cross-Sectional Studies , Female , Humans , Male , Self Report , Skin Diseases/therapy , United Kingdom
5.
Adv Exp Med Biol ; 1354: 299-314, 2022.
Article in English | MEDLINE | ID: mdl-34807448

ABSTRACT

Dairy and beef cattle make a vital contribution to global nutrition, and since their domestication, they have been continuously exposed to natural and artificial selection to improve production characteristics. The technologies of transgenesis and gene editing used in cattle are responsible for generating news characteristics in bovine breeding, such as alteration of nutritional components of milk and meat enhancing human health benefits, disease resistance decreasing production costs and offering safe products for human food, as well as the recombinant protein production of biomedical significance. Different methodologies have been used to generate transgenic cattle as bioreactors. These methods include the microinjection of vectors in pronuclear, oocyte or zygote, sperm-mediate transgenesis, and somatic cell nuclear transfer. Gene editing has been applied to eliminate unwanted genes related to human and animal health, such as allergy, infection, or disease, and to insert transgenes into specific sites in the host genome. Methodologies for the generation of genetically modified cattle are laborious and not very efficient. However, in the last 30 years, transgenic animals were produced using many biotechnological tools. The result of these modifications includes (1) the change of nutritional components, including proteins, amino acids and lipids for human nutrition; (2) the removal allergic proteins milk; (3) the production of cows resistant to disease; or (4) the production of essential proteins used in biomedicine (biomedical proteins) in milk and blood plasma. The genetic modification of cattle is a powerful tool for biotechnology. It allows for the generation of new or modified products and functionality that are not currently available in this species.


Subject(s)
Milk , Nuclear Transfer Techniques , Animals , Animals, Genetically Modified , Bioreactors , Biotechnology , Cattle , Female
8.
Res Vet Sci ; 122: 22-28, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30448391

ABSTRACT

This study investigated: 1) the kinetics of oocyte chromatin configuration during in vitro maturation (IVM) of caprine and bovine oocytes; and 2) the effect of in vitro pre-maturation (IVPM) with cilostamide with or without association of the follicular wall (FW) on the same parameters. In experiment I, cumulus-oocyte complexes (COCs) were cultured in vitro in a standard maturation medium for 6, 12, 18 or 30 h. For experiment II, the COCs were cultured for 30 h, either in a standard IVM medium or in IVPM containing cilostamide (10 or 20 µM) and FW alone or in combination, for 6 or 12 h before the onset of maturation. The MII rate was similar (P > .05) between 18 and 30 h of maturation, both of which were higher (P < .05) than 6 and 12 h IVM in both species (Experiment I). Contrary to caprine, all IVPM treatments presented a higher (P < .05) percentage of bovine oocytes arrested at the GV stage than the control treatment after 6 h of culture. The percentage of MII oocytes after 30 h (IVPM+IVM) of culture in bovine oocytes treated with 10 µM cilostamide associated with FW and FW alone cultured for 6 h presented MII percentages similar to the control. However, in caprine, these treatments significantly reduced the percentages of MII in relation to the control treatment (Experiment II). In conclusion, the combination of concentration-exposure time to cilostamide during IVPM delayed meiotic progression in bovine after 6 and 12 h of culture. However, overall the culture period (IVPM+IVM) influenced the oocyte chromatin configuration and kinetics in both species.


Subject(s)
Cattle , Goats , In Vitro Oocyte Maturation Techniques/veterinary , Oocytes/drug effects , Quinolones/pharmacology , Animals , Female , In Vitro Oocyte Maturation Techniques/methods , Kinetics , Meiosis , Oocytes/physiology
9.
Cytometry B Clin Cytom ; 94(6): 935-940, 2018 11.
Article in English | MEDLINE | ID: mdl-30350400

ABSTRACT

BACKGROUND: Bone marrow (BM) aspirate analysis by flow cytometry (FC) is a key hematopathologic technique but dry taps hinder diagnosis. We describe the utility and limitations of a method of mechanically disaggregating BM trephine biopsies for FC. METHOD: Trephine biopsies mechanically disaggregated for FC between 2010 and 2016 were reviewed. We recorded cell yield, pathological findings, and turnaround time. Where available, results of trephine FC were compared with aspirate FC. RESULTS: Eighty BM trephine samples were processed, representing 4.3% of BM biopsies. Mechanical disaggregation yielded cellular samples in 98% of cases (median yield 3 × 106 nucleated cells). The most frequent diagnoses were B cell lymphoproliferative disorders (n = 26) and acute leukemia (n = 18). Trephine FC correlated with histochemistry findings in 94% of cases, but two Hodgkin lymphoma infiltrates were missed by FC, and trephine FC underestimated B cell infiltrate burden compared to immunohistochemistry (IHC). Fluorescence intensity of CD34 and CD45 on disease populations was similar in processed trephine and contemporaneous aspirate samples. Trephine FC results were available 2 days earlier than stained IHC slides. CONCLUSION: Mechanical disaggregation of BM trephine samples provided a cellular suspension suitable for diagnostic FC in most cases. Limitations were similar to aspirate FC: disease burden was underestimated and some infiltrates were missed. Trephine FC results were available earlier than trephine IHC. We conclude that trephine FC is a useful technique to complement trephine IHC in the event of a failed aspirate, providing rapid diagnostic immunophenotyping. © 2018 International Clinical Cytometry Society.


Subject(s)
Biopsy , Bone Marrow/pathology , Flow Cytometry , Adolescent , Adult , Aged , Aged, 80 and over , Bone Marrow/immunology , Bone Marrow Examination , Female , Humans , Immunohistochemistry , Immunophenotyping , Male , Middle Aged , Retrospective Studies , Young Adult
10.
Haemophilia ; 24 Suppl 6: 87-94, 2018 May.
Article in English | MEDLINE | ID: mdl-29878652

ABSTRACT

INTRODUCTION: Information from the genes encoding factor VIII (F8) and IX (F9) is used in reproductive planning and to inform inhibitor formation, bleeding severity and response to therapies. Advances in technology and our understanding of the human genome now allows more comprehensive methods to study genomic variation and its impact on haemophilia. AIMS: The My Life Our Future (MLOF) programme was begun in 2012 to provide genetic analysis and to expand research in haemophilia through a research repository. METHODS: MLOF enrolled haemophilia A and B patients followed at haemophilia treatment centers in the U.S., including, since 2015, known and potential genetic carriers. Initial F8 and F9 DNA analysis was performed utilizing a next generation sequencing approach which allowed simultaneous detection of F8 inversions and other variants. Candidate variants were confirmed using a second method and multiplex ligation-dependent probe amplification was used to detect structural variants. RESULTS: The initial phase of MLOF completed enrollment in December 2017 with 11,356 patients, genetic carriers, and potential carriers enrolled. In the 9453 subjects in whom analysis is complete, 687 unique previously unreported variants were found. Simultaneous sequencing of the F8 and F9 genes resulted in identification of non-deleterious variants previously reported as causative in haemophilia. DNA from 5141 MLOF subjects has undergone whole genome sequencing through the NHLBI TOPMed programme of the U.S. NIH. CONCLUSION: MLOF has provided genetic information for patients and their families to help inform clinical care and has established a repository of data and biospecimens to further advance haemophilia research.


Subject(s)
Genotype , Hemophilia A/genetics , Phenotype , Whole Genome Sequencing/methods , Hemophilia A/diagnosis , Humans , Prognosis
11.
Theriogenology ; 107: 95-103, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29145066

ABSTRACT

The search for non-invasive signs of oocyte meiotic competence is very important for the development of in vitro follicle culture (IVFC) systems. The aims of the present study were: (1) to investigate the effect of in vitro maturation (IVM) of in vivo grown goat COCs, in group or individually, on oocyte chromatin configuration (Experiment 1), and (2) the influence of IVFC period (12 vs. 18 days) on the ability of the oocyte to resume meiosis immediately after IVFC (before in vitro maturation; IVM), or after IVM (Experiment 2). In experiment 1, in vivo grown cumulus-oocyte complexes (COCs) were submitted to IVM in groups (10 COCs/100 µL-drop) or individually (1 COC/10 µL-drop), and chromatin configuration was assessed. In experiment 2, isolated follicles were individually cultured for 12 or 18 days, and submitted to individual IVM afterwards. The following end points were evaluated: follicular growth and morphology, oocyte diameter, viability and chromatin configuration, as well as individual follicular estradiol production. Similar maturation rates were obtained between in vivo grown COCs matured individually and in groups (66.7% vs. 63.6%, respectively) (Experiment 1). Only after 18 days of IVFC, oocytes were able to grow during IVM, reaching a mean oocyte diameter of 119 µm. Also, this treatment produced the highest rate of metaphase II oocytes (46.2% out of the total number of cultured follicles). Finally, it was observed that follicles with a daily growth rate >7.1 µm/day (fast-growing) and that reached at least 600 µm in diameter, were more likely (P < 0.05) to produce oocytes capable of attaining MII. In conclusion, caprine oocytes can be individually matured in vitro, as efficiently as in groups. This result was essential to pair in vitro follicle development and in vitro oocyte maturation with specific individual follicles. Using this approach, it was possible to establish non-invasive signs for the efficiency of IVFC based on follicle daily growth rate and diameter, and oocyte diameter: follicle daily growth >7 µm, follicle diameter of at least 600 µm, and oocyte diameter ≥120 µm. In addition, 18 days seems to be the most suitable culture time for caprine early antral follicles.


Subject(s)
Goats/physiology , In Vitro Oocyte Maturation Techniques/veterinary , Meiosis/physiology , Oocytes/physiology , Ovarian Follicle/physiology , Animals , Cell Size , Chromatin , Estradiol/metabolism , Female , Oocytes/cytology
12.
Mol Hum Reprod ; 23(4): 269, 2017 04 01.
Article in English | MEDLINE | ID: mdl-28369598
13.
Theriogenology ; 90: 32-41, 2017 Mar 01.
Article in English | MEDLINE | ID: mdl-28166985

ABSTRACT

The aims of this study were: (1) to evaluate the effect of different insulin concentrations, alone or in combination with either a fixed FSH concentration or increasing FSH concentrations on the in vitro culture of isolated caprine preantral follicles and (2) to analyze the efficiency of two IVM media and maturation culture systems (with or without coculture with in vivo grown oocytes) on the meiosis resumption. Secondary follicles were cultured for 18 days in a basic medium supplemented with low- or high-insulin concentration alone or with a fixed FSH concentration or with increasing FSH concentrations. Oocytes grown in vivo or in vitro were matured alone or cocultured. The high-insulin concentration associated with fixed FSH treatment had higher meiotic resumption rate (P < 0.05) and was the only treatment capable of producing oocytes in metaphase II. The rates of germinal vesicle, germinal vesicle breakdown, metaphase I, metaphase II (MII), meiotic resumption, and oocyte diameter were similar between the maturation media. In conclusion, a basic medium supplemented with 10-µg/mL insulin and 100-µg/mL FSH throughout the culture period improved meiotic resumption rate and produced MII oocytes from caprine preantral follicles cultured in vitro. The MII rate was similar between in vivo and in vitro grown oocytes ≥110 µm.


Subject(s)
Coculture Techniques/veterinary , Follicle Stimulating Hormone/pharmacology , In Vitro Oocyte Maturation Techniques/veterinary , Insulin/pharmacology , Oocytes/growth & development , Ovarian Follicle/drug effects , Animals , Culture Media , Female , Goats , In Vitro Oocyte Maturation Techniques/methods , Meiosis
14.
Biometrika ; 104(4): 939-952, 2017 Dec.
Article in English | MEDLINE | ID: mdl-29422695

ABSTRACT

We consider shape restricted nonparametric regression on a closed set [Formula: see text], where it is reasonable to assume the function has no more than H local extrema interior to [Formula: see text]. Following a Bayesian approach we develop a nonparametric prior over a novel class of local extremum splines. This approach is shown to be consistent when modeling any continuously differentiable function within the class considered, and is used to develop methods for testing hypotheses on the shape of the curve. Sampling algorithms are developed, and the method is applied in simulation studies and data examples where the shape of the curve is of interest.

15.
Int J Obes (Lond) ; 41(1): 149-158, 2017 01.
Article in English | MEDLINE | ID: mdl-27773938

ABSTRACT

BACKGROUND: Consumption of dietary fat is one of the key factors leading to obesity. High-fat diet (HFD)-induced obesity is characterized by induction of inflammation in the hypothalamus; however, the temporal regulation of proinflammatory markers and their impact on hypothalamic appetite-regulating neuropeptide Y/agouti-related peptide (NPY/AgRP) neurons remains undefined. METHODS: Mice were injected with an acute lipid infusion for 24 h or fed a HFD over 8-20 weeks. Characterized mouse NPY/AgRP hypothalamic cell lines were used for in vitro experimentation. Immunohistochemistry in brain slices or quantitative real-time PCR in cell lines, was performed to determine changes in the expression of key inflammatory markers and neuropeptides. RESULTS: Hypothalamic inflammation, indicated by tumor necrosis factor (TNF)-α expression and astrocytosis in the arcuate nucleus, was evident following acute lipid infusion. HFD for 8 weeks suppressed TNF-α, while significantly increasing heat-shock protein 70 and ciliary neurotrophic factor, both neuroprotective components. HFD for 20 weeks induced TNF-α expression in NPY/AgRP neurons, suggesting a detrimental temporal regulatory mechanism. Using NPY/AgRP hypothalamic cell lines, we found that palmitate provoked a mixed inflammatory response on a panel of inflammatory and endoplasmic reticulum (ER) stress genes, whereas TNF-α significantly upregulated IκBα, nuclear factor (NF)-κB and interleukin-6 mRNA levels. Palmitate and TNF-α exposure predominantly induced NPY mRNA levels. Utilizing an I kappa B kinase ß (IKKß) inhibitor, we demonstrated that these effects potentially occur via the inflammatory IKKß/NF-κB pathway. CONCLUSIONS: These findings indicate that acute lipid and chronic HFD feeding in vivo, as well as acute palmitate and TNF-α exposure in vitro, induce markers of inflammation or ER stress in the hypothalamic appetite-stimulating NPY/AgRP neurons over time, which may contribute to a dramatic alteration in NPY/AgRP content or expression. Acute and chronic HFD feeding in vivo temporally regulates arcuate TNF-α expression with reactive astrocytosis, which suggests a time-dependent neurotrophic or neurotoxic role of lipids.


Subject(s)
Appetite/drug effects , Diet, High-Fat/adverse effects , Hypothalamus/pathology , Inflammation/chemically induced , Neurons/drug effects , Neuropeptide Y/metabolism , Palmitates/pharmacology , Tumor Necrosis Factor-alpha/pharmacology , Animals , Arcuate Nucleus of Hypothalamus/pathology , Disease Models, Animal , Gene Expression Regulation , Hypothalamus/drug effects , Inflammation/pathology , Interleukin-6/metabolism , Male , Mice , Mice, Inbred C57BL , Neurons/metabolism , Obesity/pathology
16.
Cereb Cortex ; 27(3): 2403-2417, 2017 Mar 01.
Article in English | MEDLINE | ID: mdl-27095824

ABSTRACT

Control of goal-directed tasks is putatively carried out via the cinguloopercular (CO) and frontoparietal (FP) systems. However, it remains unclear whether these systems show dissociable moment-to-moment processing during distinct stages of a trial. Here, we characterize dynamics in the CO and FP networks in a meta-analysis of 5 decision-making tasks using fMRI, with a specialized "slow reveal" paradigm which allows us to measure the temporal characteristics of trial responses. We find that activations in left FP, right FP, and CO systems form separate clusters, pointing to distinct roles in decision-making. Left FP shows early "accumulator-like" responses, suggesting a role in pre-decision processing. CO has a late onset and transient response linked to the decision event, suggesting a role in performance reporting. The majority of right FP regions show late onsets with prolonged responses, suggesting a role in post-recognition processing. These findings expand upon past models, arguing that the CO and FP systems relate to distinct stages of processing within a trial. Furthermore, the findings provide evidence for a heterogeneous profile in the FP network, with left and right FP taking on specialized roles. This evidence informs our understanding of how distinct control networks may coordinate moment-to-moment components of complex actions.


Subject(s)
Brain/physiology , Decision Making/physiology , Brain Mapping , Executive Function/physiology , Humans , Magnetic Resonance Imaging , Neural Pathways/physiology
17.
Reprod Fertil Dev ; 29(6): 1144-1154, 2017 Jun.
Article in English | MEDLINE | ID: mdl-27166082

ABSTRACT

The aim of the present study was to evaluate the effect of anti-Müllerian hormone (AMH), with and without FSH, on the in vitro development of isolated caprine preantral follicles, as well as follicular steroid production and mRNA levels of AMH, hormone receptors (AMH and FSH), CYP19A1 (cytochrome P450, family 19, subfamily A, polypeptide 1), CYP17 (cytochrome P450, family 17, subfamily A, polypeptide 1), HSD3B (3-beta-hydroxysteroid dehydrogenase) and Myc (myelocytomatosis oncogene). Isolated secondary follicles were cultured in minimum essential medium alpha (α-MEM+) alone or supplemented with 50ng mL-1 AMH and/or 100ng mL-1 FSH added sequentially on different days of culture. Follicles were cultured for a total of 18 days, with different media during the first (Days 0-9) and second (Days 10-18) halves of the culture period, resulting in six treatment groups, as follows: α-MEM+/α-MEM+, FSH/FSH, AMH/AMH, AMH+FSH/AMH+FSH, AMH/FSH, and FSH/AMH. Follicle development was evaluated on the basis of follicular growth, oocyte maturation and steroid secretion. There was a decrease in follicular growth rate in the AMH, AMH+FSH and AMH/FSH treatment groups compared with α-MEM+ and FSH treatment groups (P<0.05). However, the different culture conditions had no effect on rates of meiotic resumption and steroid secretion (P>0.05). Moreover, follicles cultured in the presence of FSH had lower levels of AMH receptor type II (AMHRII) mRNA compared with non-cultured control (freshly isolated follicles), and the AMH and AMH/FSH treatment groups. In conclusion, AMH reduces the follicular growth rate of isolated goat preantral follicles in vitro without affecting follicular survival.


Subject(s)
Anti-Mullerian Hormone/metabolism , Follicle Stimulating Hormone/metabolism , Gene Expression Regulation, Developmental , Oogenesis , Ovarian Follicle/metabolism , Receptors, FSH/agonists , Receptors, Peptide/agonists , Receptors, Transforming Growth Factor beta/agonists , Abattoirs , Animals , Anti-Mullerian Hormone/genetics , Anti-Mullerian Hormone/pharmacology , Brazil , Cattle , Cell Proliferation/drug effects , Cell Size/drug effects , Cell Survival/drug effects , Crosses, Genetic , Estradiol/metabolism , Female , Follicle Stimulating Hormone/genetics , Follicle Stimulating Hormone/pharmacology , Gene Expression Regulation, Developmental/drug effects , Goats , Humans , Oogenesis/drug effects , Ovarian Follicle/cytology , Ovarian Follicle/drug effects , Progesterone/metabolism , Receptors, FSH/genetics , Receptors, FSH/metabolism , Receptors, Peptide/genetics , Receptors, Peptide/metabolism , Receptors, Transforming Growth Factor beta/genetics , Receptors, Transforming Growth Factor beta/metabolism , Recombinant Proteins/metabolism , Recombinant Proteins/pharmacology , Testosterone/metabolism , Tissue Culture Techniques
18.
Mol Hum Reprod ; 23(4): 248-256, 2017 04 01.
Article in English | MEDLINE | ID: mdl-27418669

ABSTRACT

The in vitro production of livestock embryos is central to several areas of animal biotechnology. Further, the use of in vitro embryo manipulation is expanding as new applications emerge. ARTs find direct applications in increasing genetic quality of livestock, producing transgenic animals, cloning, artificial insemination, reducing disease transmission, preserving endangered germplasm, producing chimeric animals for disease research, and treating infertility. Whereas new techniques such as nuclear transfer and intracytoplasmic sperm injection are now commonly used, basic embryo culture procedures remain the limiting step to the development of these techniques. Research over the past 2 decades focusing on improving the culture medium has greatly improved in vitro development of embryos. However, cleavage rates and viability of these embryos is reduced compared with in vivo indicating that present in vitro systems are still not optimal. Furthermore, the methods of handling mammalian oocytes and embryos have changed little in recent decades. While pipetting techniques have served embryology well in the past, advanced handling and manipulation technologies will be required to efficiently implement and commercialize the basic biological advances made in recent years. Microfluidic systems can be used to handle gametes, mature oocytes, culture embryos, and perform other basic procedures in a microenvironment that more closely mimic in vivo conditions. The use of microfluidic technologies to fabricate microscale devices has being investigated to overcome this obstacle. In this review, we summarize the development and testing of microfabricated fluidic systems with feature sizes similar to the diameter of an embryo for in vitro production of pre-implantation mammalian embryos.


Subject(s)
Embryo Culture Techniques/methods , Embryonic Development/physiology , Microfluidic Analytical Techniques/methods , Microfluidics/methods , Animals , Animals, Genetically Modified , Conservation of Natural Resources/methods , Culture Media/chemistry , Culture Media/pharmacology , Embryo, Mammalian , Embryonic Development/drug effects , Female , Livestock , Mice , Microfluidics/instrumentation , Nuclear Transfer Techniques/instrumentation , Sperm Injections, Intracytoplasmic/instrumentation , Sperm Injections, Intracytoplasmic/methods
19.
Br Dent J ; 221(1): 31-6, 2016 Jul 08.
Article in English | MEDLINE | ID: mdl-27388088

ABSTRACT

Objective This pilot study aimed to produce and evaluate training resources and training in oral health care, including oral hygiene, for carers in care homes in Surrey and Medway.Methods During two training days, for carers from these homes, short, interactive presentations were given on a range of topics relevant to oral health care and oral hygiene of older people, followed by practical training. Prior to any training all attendees completed a 39 question questionnaire to establish their baseline knowledge of oral health and hygiene. At the end of the training day they completed an evaluation form. Fourteen weeks later, they were visited at their place of work and completed the same questionnaire again. Differences in responses between baseline and after 14 weeks were statistically tested using the chi-squared test.Results Sixty-six carers attended the training sessions and 44 were followed up 14 weeks later. The results showed an improvement in carer knowledge at follow up. The majority of carers (36/44) spoke English as their first language. They had a mean age of 41 years, 37 were female and 7 male. They had worked as carers for a mean of 10.9 years (range 4 months-34 years). Over 90% stated that the training day fully met or exceeded their requirements and expectations.Conclusions The results indicated improvements in carer knowledge. However, the carers were atypical of carers in general, as they were self-selected and well-motivated. Nevertheless the content of the training day and the questionnaire should inform future work in this area.


Subject(s)
Caregivers , Oral Health , Adult , Education , Female , Health Knowledge, Attitudes, Practice , Humans , Male , Mouth , Pilot Projects , Surveys and Questionnaires
20.
Anal Methods ; 8(14): 2880-2886, 2016 Apr 14.
Article in English | MEDLINE | ID: mdl-37457919

ABSTRACT

Sample preparation, including bacterial lysis, remains a hurdle in the realization of complete point-of-care tests for many pathogens. Here, we developed a sample preparation methodology for enzymatic lysis and sample heating for low-resource, point-of-care applications. We show an instrument-free chemical heater system for rapid lysis of a gram-positive bacterium (Staphylococcus aureus) and an RNA virus (human respiratory syncytial virus) using a dried lysis enzyme mixture (achromopeptidase) for S. aureus. After a lysis step (<1 minute), lysis enzymes are heat deactivated (<5 minutes) using a simple disposable chemical heater. We demonstrated that both DNA and RNA in the heat-treated sample could be directly amplified without purification, even in the presence of a clinically-obtained human nasal sample. This simple approach to dry enzyme storage and sample heating is adaptable to many applications where samples need to be lysed, including use in low-resource laboratories and in single-use or cartridge-based point-of-care diagnostic devices.

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