Use of gene profiling to evaluate the effects of a feed additive on immune function in periparturient dairy cattle.

Objectives were to investigate mechanisms by which a nutritional supplement alters immunity in dairy cattle. Our hypothesis was that feeding this product to dairy cattle altered neutrophil gene expression. Eight periparturient Jersey cattle were randomly assigned to one of two treatments: control and treated. Control animals were fed a dry cow ration for 1 month prior to calving. The treated cows were fed the same ration supplemented with OmniGen-AF. Following calving, blood samples were taken and neutrophils were prepared after which RNA was extracted. Gene expression in neutrophils of treated versus control-fed animals was then assessed using bovine-total leukocyte (BOTL-5) arrays. Eighteen genes were differentially regulated in the experimental group and of these, twice as many were up-regulated as down-regulated. Patterns of changes indicated that the additive might alter neutrophil apoptosis, signaling and sensitivity. Two of the regulated genes [interleukin-1beta converting enzyme (ICE) and interleukin-4 receptor (IL-4R)] were investigated in more detail using quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR). Each was found to be elevated by the feeding of experimental product. Increased expression of ICE indicates potential for enhanced neutrophil expression of interleukin-1beta (IL-1beta), a cytokine which plays roles in the inflammatory response and which stimulates adaptive immunity following innate immune activation. Altered expression of IL-4R indicates potential for changes in neutrophil apoptosis. The experiment identified mechanisms by which the additive altered neutrophil gene expression. While many nutrients support the immune system, we have shown that a non-traditional nutritional approach may also have utility in modulating immune function.

Post-mortem microbiology in sudden death: sampling protocols proposed in different clinical settings.

BACKGROUND: Autopsies, including minimally invasive autopsies, are a powerful tool for determination of the cause of death. When a patient dies from an infection, microbiology is crucial to identify the causative organism. Post-mortem microbiology (PMM) aims to detect unexpected infections causing sudden deaths; confirm clinically suspected but unproven infection; evaluate the efficacy of antimicrobial therapy; identify emergent pathogens; and recognize medical errors. Additionally, the analysis of the thanatomicrobiome may help to estimate the post-mortem interval. AIMS: The aim was to provide advice in the collection of PMM samples and to propose sampling guidelines for microbiologists advising autopsy pathologists facing different sudden death scenarios. SOURCES: A multidisciplinary team with experts in various fields of microbiology and autopsies on behalf of the ESGFOR (ESCMID - European Society of Clinical Microbiology and Infectious Diseases - study group of forensic and post-mortem microbiology and in collaboration with the European Society of Pathology) developed this narrative review based on a literature search using MedLine and Scopus electronic databases supplemented with their own expertise. CONTENT: These guidelines address measures to prevent sample contamination in autopsy microbiology; general PMM sampling technique; protocols for PMM sampling in different scenarios and using minimally invasive autopsy; and potential use of the evolving post-mortem microbiome to estimate the post-mortem interval. IMPLICATIONS: Adequate sampling is paramount to identify the causative organism. Meaningful interpretation of PMM results requires careful evaluation in the context of clinical history, macroscopic and histological findings. Networking and closer collaboration among microbiologists and autopsy pathologists is vital to maximize the yield of PMM.

Transportation stress alters the circulating steroid environment and neutrophil gene expression in beef bulls.

Stress and its association with altered immune function and incidence of respiratory diseases in cattle have lead to concerns over animal health and welfare during truck transportation. Previously, bulls subjected to transportation stress displayed altered expression of candidate neutrophil genes, warranting a broader investigation of the neutrophil transcriptome and possible associations with fluctuations in circulating steroid hormones. In the current study, blood was collected from six Belgian BluexFriesian bulls at -24, 0, 4.5, 9.75, 14.25, 24, and 48h relative to initiation of 9h of truck transportation. Plasma concentrations of cortisol, dehydroepiandrosterone (DHEA), progesterone, and testosterone were measured; cortisol:DHEA ratios were computed. Neutrophil gene expression was monitored by microarray analysis using bovine immunobiology (BOTL-5) microarrays. Eighty-eight genes were identified as being differentially expressed at P<0.05. Putatively affected genes were grouped into ontological clusters; those of greatest interest for qRT-PCR validation were involved in immune response, apoptosis, wound healing, and several of currently unknown function. Confirmed gene expression changes supported the dramatic effects of transportation stress on the bovine neutrophil transcriptome. Temporal correlations between gene expression profiles and circulating total leukocyte and neutrophil counts were apparent. However, few relationships between gene expression and plasma steroid profiles were detected, possibly due to the biological time-lag between these variables not captured by the blood collection schedule. Further investigation into the factors underlying neutrophil gene expression changes and validations at the protein and cell behavior levels will lead to a better understanding of altered innate immunity in cattle during transportation stress.

Depressed DHEA and increased sickness response behaviors in lame dairy cows with inflammatory foot lesions.

Lameness is a multifactorial condition influenced by the environment, genetics, management and nutrition. Detection of lameness is subjective and currently limited to visual locomotion observations which lack reliability and sensitivity. The objective of this study was to search for potential biomarkers of inflammatory foot lesions that underlie most cases of lameness in dairy cows, with a focus on the sickness response and relevant endocrine, immune and behavioral changes. Serum and peripheral blood mononuclear cells (PBMC) were collected from eight sound and eight lame high-producing Holstein cows. Immune cell activation was investigated in PBMCs using a candidate gene approach in which the expression of pro-opiomelanocortin, interleukin-1beta, l-selectin, matrix metalloproteinase-9 and glucocorticoid receptor-alpha was measured via quantitative real time-RT-PCR. Endocrine changes were investigated by monitoring serum concentrations of cortisol and dehydroepiandrosterone (DHEA). Additionally, systematic behavioral observations were carried out to characterize a behavioral profile associated with a sickness response typical of this condition. Lame cows showed significantly lower eating (P=0.01) and ruminating (P=0.01) behaviors and higher incidence of self-grooming (P=0.04) compared to sound cows. Lame cows also showed a 23% decrease in serum DHEA (P=0.01) and 65% higher cortisol:DHEA ratio (P=0.06) compared to sound cows. However, no significant differences were found in candidate gene expression between lame and sound cows. In association with sickness behaviors, serum DHEA concentration and cortisol:DHEA ratio are promising objective indicators of inflammatory foot lesions in dairy cattle and may be useful as diagnostic targets for animals in need of treatment.

Hsl1p, a Swe1p inhibitor, is degraded via the anaphase-promoting complex.

Ubiquitination and subsequent degradation of critical cell cycle regulators is a key mechanism exploited by the cell to ensure an irreversible progression of cell cycle events. The anaphase-promoting complex (APC) is a ubiquitin ligase that targets proteins for degradation by the 26S proteasome. Here we identify the Hsl1p protein kinase as an APC substrate that interacts with Cdc20p and Cdh1p, proteins that mediate APC ubiquitination of protein substrates. Hsl1p is absent in G(1), accumulates as cells begin to bud, and disappears in late mitosis. Hsl1p is stabilized by mutations in CDH1 and CDC23, both of which result in compromised APC activity. Unlike Hsl1p, Gin4p and Kcc4p, protein kinases that have sequence homology to Hsl1p, were stable in G(1)-arrested cells containing active APC. Mutation of a destruction box motif within Hsl1p (Hsl1p(db-mut)) stabilized Hsl1p. Interestingly, this mutation also disrupted the Hsl1p-Cdc20p interaction and reduced the association between Hsl1p and Cdh1p in coimmunoprecipitation studies. These findings suggest that the destruction box motif is required for Cdc20p and, to a lesser extent, for Cdh1p to target Hsl1p to the APC for ubiquitination. Hsl1p has been previously shown to inhibit Swe1p, a protein kinase that negatively regulates the cyclin-dependent kinase Cdc28p, by promoting Swe1p degradation via SCF(Met30) in a bud morphogenesis checkpoint. Results of the present work indicate that Hsl1p is degraded in an APC-dependent manner and suggest a link between the SCF (Skp1-cullin-F box) and APC-proteolytic systems that may help to coordinate the proper progression of cell cycle events.

Transportation stress in young bulls alters expression of neutrophil genes important for the regulation of apoptosis, tissue remodeling, margination, and anti-bacterial function.

The transportation of beef cattle results in a stress response that is associated with increased susceptibility and severity of respiratory diseases, presumably due to an alteration in immune function. Neutrophils are phagocytic immune cells important in lung defense and are also targets of the stress response. The objective of this study was to determine if a 9h transportation of young bulls by road induced changes in the expression of candidate genes known to be important in neutrophil-mediated defense and inflammation in the lung. These neutrophil genes encompassed functions of apoptosis (A1 and Fas), tissue remodeling (MMP-9), vascular margination (L-selectin), bacterial killing (BPI), and wound healing (betaglycan), as well as responsiveness of the cells to stress-induced increases in glucocorticoid hormones (GRalpha). To explore gene expression changes, blood was collected, plasma harvested, and neutrophils isolated from six Belgian Blue x Friesian bulls (231+/-7.0 kg in weight; 282+/-4 days of age) at -24, 0, 4.5, 9.75, 14.25, 24, and 48h relative to commencement of a 9h road transportation by truck. Plasma cortisol concentrations were elevated at 4.5 and 9.75h, peaking at 50.64+/-4.46 ng/mL (P<0.0001) and confirming that the animals experienced stress. Blood neutrophil count was elevated between 4.5 and 14.25h (P<0.0001), reaching a peak that was over 3-fold higher than the -24h concentration. Neutrophil Fas gene expression was acutely down-regulated (P=0.02) by transportation stress, while expressions of MMP-9, l-selectin, and BPI were profoundly up-regulated (P=0.003, 0.002, and <0.001 respectively). However, no changes in neutrophil expressions of betaglycan, GRalpha, and A1 were detected. It is concluded that a 9h transportation of young bulls induces a gene expression signature in blood neutrophils that increases their circulating numbers and may enhance their pro-inflammatory and anti-bacterial potential.

Analysis of the bovine neutrophil transcriptome during glucocorticoid treatment.

The objective of this study was to characterize a large portion of the bovine neutrophil transcriptome following treatment with the anti-inflammatory glucocorticoid dexamethasone (Dex). Total RNA was isolated from blood neutrophils of healthy cattle (5 castrated male Holsteins) immediately following cell purification (0 h) or after ex vivo aging for 4 h with or without added Dex. Additional neutrophils were cotreated with a glucocorticoid receptor (GR) antagonist (RU486) and Dex for 4 h. RNA was amplified, dye labeled (Cy3 or Cy5), and hybridized to a series of National Bovine Functional Genomics Consortium (NBFGC) microarrays. LOWESS data normalization followed by mixture model analyses showed that 11.15% of the spotted NBFGC cDNAs (2,036/18,263) were expressed in 4-h (untreated) neutrophils. Subsequent two-step mixed-model analysis detected (P < or = 0.05) 1,109 differentially expressed genes, of which contrast analysis indicated those that were independently responsive to aging (1,064), Dex (502), RU486 + Dex (141), or RU486 (357). In silico analysis revealed that 416 of the differentially expressed genes are unknown, 59 did not cluster well based on known function, and 634 clustered into 20 ontological categories. Independent validation of differential expression was done for 14 of the putatively Dex-responsive genes across these categories. Results showed that Dex induced rapid translocation of GR into the neutrophil nucleus and signaled dramatic alterations in expression of genes that delay apoptosis, enhance bactericidal activity, and promote tissue remodeling without inflammation or fibrosis. Thus these findings revealed hitherto unappreciated plasticity of blood neutrophils and potentially novel anti-inflammatory/wound-healing actions of glucocorticoids.

Short communication: A potential antiapoptotic phenotype in neutrophils of cows milked once daily in early lactation.

The objectives of this experiment were to compare the circulating concentrations of cortisol and determine whether these correlated with the expression profiles of a set of candidate apoptosis genes in neutrophils of Holstein-Friesian cows milked once vs. 3 times daily for 28 d postpartum. Cows on the once-daily milking regimen had significantly higher plasma cortisol concentrations on d 3, 14, and 28 postpartum than did those milked 3 times daily. On d 3 postpartum, when differences in cortisol and neutrophil counts were highest between the groups, mean mRNA abundance of nuclear factor kappaB p65 subunit, IkappaBalpha, X-linked inhibitor of apoptosis; and heat shock protein 70 were higher in neutrophils of the cows milked once daily than in cows milked 3 times daily. However, no correlations were detected among plasma cortisol concentration, neutrophil count, or neutrophil gene expression in this study. Results suggest that the modest neutrophilia associated with once daily milking of cows immediately postpartum may be related to modifications in the cells' apoptotic program by factors other than cortisol.

Effects of dexamethasone on bovine circulating T lymphocyte populations.

In cattle, gamma delta T cells represent a higher proportion of circulating T cells than in humans. Bovine gamma delta T cells can be recognized by expression of gamma delta cell receptor (gamma delta TCR) determinants or by a 215/230-kDa surface antigen (WC1). WC1 is expressed on 90% or more of circulating bovine gamma delta T cells. The effects of dexamethasone on this and other subsets (CD3, CD2, CD4, CD8) of peripheral blood T lymphocytes were determined by flow cytometric analysis. Twelve 15-month old bulls were injected with dexamethasone (0.04 mg/kg/day) for 3 consecutive days and four bulls were untreated controls. Blood samples were collected daily for 3 days before dexamethasone injections and for an additional 7 days starting on the third day. Data were recorded as percent positive cells and as mean fluorescent intensity (MFI) of positive cells. Initially, CD3+ cells represented 65-73% of all peripheral blood clear cells (PBMC). Dexamethasone reduced CD3+/- cells (PBMC). Dexamethasone reduced CD3+ cells to 30% and these recovered to 50% positive cells by 9 days after the last dexamethasone injection. Loss of CD3+ cells was not due to reductions in alpha beta T cells because dexamethasone did not influence the percent CD2+, CD4+, or CD8+ cells. However, percent WC1+ cells rapidly declined from a baseline of 26.4% of PBMC to < 6% by the final injection. During injections, the MFI of WC1 increased. The MFI of WC1 returned to control values 7 days after the last injection of dexamethasone, but the percent gamma delta T cells recovered to only 14% WC1+ PBMC by the final day of the study. During its maximum effects on WC1, dexamethasone also caused a profound decrease of L-selectin MFI on remaining PBMC (mostly alpha beta T cells and monocyte/macrophages). In a second trial, two-color analyses determined that dexamethasone did not increase apoptosis in WC1+ cells and did not reduce L-selectin MFI on either CD2+ of WC1+ cells. The cumulative results suggested that dexamethasone promoted gamma delta T cell migration out of peripheral blood via an L-selectin-independent mechanism and that dexamethasone did not alter alpha beta T cell migration kinetics.

Regulation of L-selectin and CD18 on bovine neutrophils by glucocorticoids: effects of cortisol and dexamethasone.

The responsiveness of bovine neutrophil L-selectin and CD18 to in vivo glucocorticoid administration was characterized by flow cytometric analysis. Blood was sampled intensively from dairy cows treated for 3 days with placebo, cortisol, or dexamethasone. Immunostaining was performed on whole blood (100 microliters) that was left unstimulated or was stimulated with platelet-activating factor (PAF; 1 microgram/ml blood) prior to incubation with fluorescein isothiocyanate-conjugated monoclonal antibodies against L-selectin and CD18. Results were expressed as the percentage of positive-staining cells and as mean fluorescence intensity (MFI) of those cells. Total leukocyte count and leukocyte differentials were also performed on all blood samples. Dexamethasone caused nearly complete down-regulation of L-selectin (P < .01) on the surface of gated cells and reduced to half the MFI of CD18 (P < .01). Compared with values for the placebo group, dexamethasone began to cause L-selectin down-regulation within 8 h after the first injection and these effects persisted until 48 h after the third injection. This was correlated in time with an acute reduction in the proportion of cells that stained positive for L-selectin (from 98% before dexamethasone injections to a low of 17% by 40 h after the first injection). Dexamethasone also caused leukocytosis and neutrophilia during this time interval. In contrast, CD18 down-regulation was delayed until 16 h after the second dexamethasone injection and persisted for roughly 8 days. However, at no time during the experiment did dexamethasone influence the proportion of gated cells staining positive for CD18 (always 100%). Effects of cortisol were generally similar in pattern to those of dexamethasone but were more subtle and more readily detected when PAF was added to blood prior to immunostaining. These results strongly suggest that one mechanism of the anti-inflammatory action of glucocorticoids is to induce dramatic down-regulation of L-selectin and CD18 adhesion molecules on blood neutrophils.

Altered insulin-like growth factor-II (IGF-II) level and IGF-binding protein-3 (IGFBP-3) protease activity in interstitial fluid taken from the skin lesion of psoriasis.

In the present study, we have investigated insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) in serum and artificially raised blister fluid from uninvolved and involved areas of nine patients with psoriasis. Both levels of IGFs and IGFBP-3, and profiles of IGFBP in serum and fluid from the uninvolved areas of these patients were comparable to those seen in normal subjects. In fluid from the involved areas, the IGF-II but not IGF-I level was significantly elevated. Among five molecular forms of IGFBP, the density of 41.5- and 38.5-kDa forms of IGFBP-3 were apparently increased in fluid from the involved areas, shown by Western ligand blotting. Radioimmunoassay further showed that the IGFBP-3 concentration in the involved areas was significantly raised. Immunoblotting revealed that the predominant form of IGFBP-3 in fluid from the uninvolved areas was a 29-kDa proteolytically modified product. In contrast, intact doublet IGFBP-3 was the main form of IGFBP-3 in fluid from the involved areas. Fluid from the involved areas but not the matched serum concentration-dependently inhibited the degradation of 125I-labeled nonglycosylated IGFBP-3 (ngIGFBP-3) caused by fluid from the uninvolved areas, suggesting the presence of an IGFBP-3 protease inhibitor(s) in psoriatic skin lesion. These findings suggest that the alterations in IGF/IGFBP system may contribute to the pathogenesis of psoriasis.

Proteolysis of insulin-like growth factor-binding protein-3 by human skin keratinocytes in culture in comparison to that in skin interstitial fluid: the role and regulation of components of the plasmin system.

Proteolysis of insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3) is an important determinant of IGF action on cells. We have investigated this in a human skin keratinocyte cell line HaCaT. Although these cells did not normally produce an active IGFBP-3 protease, addition of plasminogen resulted in a dose-dependent proteolysis of endogenous and exogenous IGFBP-3, producing fragments similar to those cleaved by skin interstitial fluid, but different from those generated by plasmin. Protease inhibitor profiles suggested the enzyme in the conditioned medium to be a calcium-dependent serine protease. Exogenous IGFBP-3 either inhibited or slightly stimulated IGF-I-induced cell proliferation when it was coincubated or preincubated with the cells, respectively. Both effects were attenuated in the presence of plasminogen. Preincubation of cells with IGF-I or long R3 IGF-I divergently changed plasminogen activator inhibitor-1 and -2 secretion, but only IGF-I blocked IGFBP-3 proteolysis. Such inhibition was also observed in a cell-free protease assay. IGF-I, however, had no effect on plasmin-induced IGFBP-3 degradation. Together, these data indicate that an IGFBP-3 protease similar to that in skin interstitial fluid is generated in plasminogen-treated HaCaT cells, and it attenuates the effects of IGFBP-3 on IGF action. IGF-I, probably by coupling with IGFBP-3, can protect it from the action of this protease.

Insulin-like growth factors (IGFs) and IGF-binding proteins in human skin interstitial fluid.

Despite extensive investigation of the insulin-like growth factor (IGF)/IGF-binding protein (IGFBP) system in the circulation and body fluids, there is no information on this in interstitial fluid. We have compared the IGF/IGFBP system in the circulation with that in fluid obtained from blisters artificially raised by negative pressure in 10 healthy volunteers. IGFBP-1, -2, -3, and -4 were all found in blister fluid, but in concentrations much lower than those in matched serum. The IGF-I, IGF-II, and IGFBP-3 levels measured by RIA were 18%, 14%, and 16% of those in serum, respectively. Fast protein liquid chromoatography showed that both IGF-I and IGFBP-3 in 150- and 50-kilodalton complexes were approximately 13% and 37%, respectively, of the corresponding peaks found in matched serum. Compared to that in serum, the IGFBP-3 in the blister fluid was predominantly in a modified 29-kilodalton form, and there was increased activity of an IGFBP-3 protease. Therefore, although IGF concentrations are much lower in interstitial fluid than in the circulation, a greater proportion of this IGF is in forms more readily available for interaction with tissue receptors. The blister fluid appears to represent physiological interstitial fluid and may provide a model for studying the physiology and pathophysiology of growth factors in the interstitial environment.

The effect of chlorpromazine on the secretion of immunoreactive beta-MSH and prolactin in man.

The effect of chlorpromazine (50 mg. im) on the plasma concentration of immunoreactive beta-melanocyte-stimulating hormone (beta-MSH) and prolactin was studied in 8 hospitalized subjects with non-endocrine skin disorders. Plasma beta-MSH concentrations remained unchanged over a period of 7 h in 6 subjects. In the remaining 2 subjects there was a slight increase. Plasma prolactin concentrations were greatly increased in all subjects 1 1/2-3 h after the injection and had almost returned to pre-injection levels by 7 h. This suggests that the control of beta-MSH secretion in man, unlike that of prolactin in man and MSH peptides in other mammals, is not predominantly inhibitory. The reason for this discrepancy may be that beta-MSH is not a natural MSH in man and occurs as part of the lipotropic hormone (LPH) or as a breakdown product.

Health and safety at necropsy.

The postmortem room is a source of potential hazards and risks, not only to the pathologist and anatomical pathology technician, but also to visitors to the mortuary and those handling the body after necropsy. Postmortem staff have a legal responsibility to make themselves aware of, and to minimise, these dangers. This review focuses specifically on those hazards and risks associated with the necropsy of infected patients, with foreign objects present in the body, and with bodies that have been contaminated by chemicals or radioactive sources.

Medical educators' personal attitudes towards the necropsy.

AIMS: To explore the personal attitudes of medical educators to the necropsy. METHODS: A "theoretical sample" of individuals, with widely disparate views, was selected from University of Sheffield Medical School affiliated staff. These individuals underwent semi-structured interviews (in private) to elucidate their personal attitudes towards the necropsy (in relation to the possibility of their own necropsy or that of a close relative). RESULTS: Nine themes pertaining to personal attitudes towards the necropsy were identified. In general medical educators had a positive attitude towards the necropsy. CONCLUSIONS: This study suggests that medical educators still believe that the necropsy is of value in medical education, despite dramatic curricular revisions in the UK, declining necropsy rates, and adverse media attention to the necropsy.

Are clinicians failing to supply adequate information when requesting a histopathological investigation?

AIMS: There is a perception among histopathologists that specimens are often received without adequate clinical details. This is the first study to determine the adequacy of information provided when histopathological investigations are requested. METHODS: Two thousand sequential requests for histological examination were assessed for adequacy and completeness. RESULTS: There was no significant difference in the demographic details supplied by physicians and surgeons. Clinical details were inadequate in 6.1% of cases: those from physicians were significantly more often adequate (98.7% v 90.6%) and more often included a diagnosis (74.4% v 38.8%) than those from surgeons. Physicians were more likely to supply their name and contact number but requests frequently lacked details of the sender. CONCLUSIONS: Specimens are infrequently received with inadequate demographic details, but clinical details and details of the sender are more often lacking. Education of clinical colleagues is required if pathologists are to manage the demand for the service.

The effect of phytoestrogens on the female genital tract.

Environmental oestrogens have been implicated in the pathogenesis of hormonally treated cancers (such as breast and prostate cancer), male infertility, and abnormalities of the male and female reproductive tracts. They may be derived from plants (phytoestrogens), pharmaceuticals, or other synthetic compounds not originally intended to have oestrogenic activity (including soy based infant formulas). This review will discuss the evidence from both animal studies and humans for an effect of these ubiquitous compounds on the development of the human female genital tract, in addition to prolonging the menstrual cycle, alleviating symptoms of the menopause, and protecting against the development of endometrial carcinoma.

Necropsy practice after the "organ retention scandal": requests, performance, and tissue retention.

AIMS: After the so called "organ retention scandal" in the UK this study set out to assess the impact on death certification and hospital (consent) necropsies, including the postmortem retention of tissues and organs. METHODS: Data were prospectively gathered over a one year period for all deaths occurring at the Royal Hallamshire Hospital, Sheffield, UK to determine the frequencies with which death certificates were completed and necropsies were requested. The seniority of the clinician undertaking these duties was recorded. Pathologists were asked to record the extent of every necropsy during the study period. The type and planned uses of tissues retained were recorded. RESULTS: Death certificates were issued for 88.5% of the 966 deaths for which clinicians completed proformas. Of these, 88.9% were issued by preregistration and senior house officers. Consent was sought for a necropsy in 6.2% of cases (usually by non-consultant staff) and was granted in 43.4% of these. The overall, medicolegal, and hospital necropsy rates were 13.4%, 9.9%, and 3.5%, respectively. Tissues were retained from 55.4% of necropsies for diagnostic purposes, although sampling does not appear to be systematic. CONCLUSIONS: Death certification and seeking consent for a necropsy are frequently delegated to junior clinical staff. This may explain the low standard of death certification reported by others and the low necropsy rate. The decline in the necropsy rate and the low rate of sampling for histological examination highlight the decline of the hospital necropsy and the lack of a systematic approach to tissue sampling.

Videos have a role in postgraduate necropsy education.

AIMS: This is the first study to investigate the usefulness of structured, scripted videos as an adjunct to the mortuary based training of histopathology trainees in necropsy techniques. METHODS: Four structured and scripted videos describing aspects of necropsy health and safety, evisceration, general dissection techniques, specialist dissection techniques, and reconstruction were shown to histopathology trainees attending the 2001 University of Sheffield short course on the autopsy. Delegates who agreed to participate in the study were asked to complete a short questionnaire seeking Likert-type and free text responses concerning the usefulness of the videos in postgraduate necropsy training. Free text responses were analysed using a themed content analysis. RESULTS: All 38 delegates who viewed the videos agreed to participate in the study. Of these, 35 found the videos enjoyable and 34 found them interesting. Thirty one felt the videos enhanced their learning experience. Advantages of the videos included the ability to learn about specialist techniques rarely encountered in the mortuary, the ability to teach large numbers of students at once, allowing students to learn at their own pace, and as a tool for revision. Repetition between the videos, a lack of interactivity, and a lack of sufficient detail on general necropsy techniques were felt by participants to be the principal disadvantages of this teaching tool. CONCLUSIONS: Videos are an acceptable teaching tool for students. They have a valuable role to play as an adjunct to dissection in teaching junior histopathology trainees about specialist necropsy dissection techniques.