Accuracy of a history of blood donation from surrogate witnesses: data from the UK TMER study.

Look-back studies of blood transfusion in Creutzfeldt-Jakob disease commonly rely on reported history from surrogate witnesses. Data from the UK Transfusion Medicine Epidemiology Review have been analysed to determine the accuracy of the blood donation history provided by the relatives of cases. Our results show that only a small percentage of cases were found to be registered as donors on UK Blood Service (UKBS) databases when there was no family report of blood donation. In contrast, a history of reported donation was less accurate.

Creutzfeldt-Jakob disease and blood transfusion: updated results of the UK Transfusion Medicine Epidemiology Review Study.

BACKGROUND AND OBJECTIVES: This paper reports the results to 31 May 2015 of an ongoing UK study to look for additional cases of variant Creutzfeldt-Jakob disease (vCJD) transmission by blood transfusion, and to seek evidence whether other subtypes of Creutzfeldt-Jakob disease (CJD) may be transmissible via blood components. MATERIALS AND METHODS: All vCJD cases of appropriate age and any sporadic CJD (sCJD) or familial CJD (fCJD) cases with a history of blood donation or transfusion are notified to the UKBS. Donation records are sought and the usage of all donations is determined by look back. Death certificates are obtained for all donors to patients with CJD and recipients of transfused components from patients with CJD who are deceased. RESULTS: The study identified 29 sCJD blood donors, of 370 reported, with transfusion to 211 recipients. Five of these recipients were reported to have died with or of dementia, but were not believed to be cases of CJD. The vCJD arm found 18 vCJD blood donors who had donated blood which was issued for clinical usage, of 24 traced donors from 177 UK vCJD cases. To date, 3 cases of vCJD have occurred in 67 recipients identified in this recipient group, and one recipient had post-mortem confirmation of abnormal prion protein deposition in the spleen (all previously reported). CONCLUSION: The results of the ongoing TMER study show no new cases of transfusion-associated vCJD since 2007 and no evidence of transfusion transmission of sCJD.

Ten-year follow-up of two cohorts with an increased risk of variant CJD: donors to individuals who later developed variant CJD and other recipients of these at-risk donors.

BACKGROUND: Transmission of variant Creutzfeldt-Jakob disease (vCJD) through blood transfusion is implicated in three deaths and one asymptomatic infection. Based on this evidence, individuals assessed to be at increased risk of vCJD through donating blood transfused to individuals who later developed vCJD, or through being other recipients of such donors, are followed up to further understand the risks of vCJD transmission through blood. OBJECTIVES: To provide a ten-year follow-up of these at-risk cohorts. METHODS: Blood donors to patients who later died from vCJD were identified by the Transfusion Medicine Epidemiological Review (TMER) study. A reverse risk probability assessment quantified the risk of blood transfusion or exposure through diet as the source of vCJD in the recipients. Donors to these recipients, and these donors' other recipients, with a probability risk above 1%, are classified as at increased risk of vCJD for public health purposes. These cohorts are monitored for any vCJD occurrences. RESULTS: A total of 112 donors and 33 other recipients of their donated blood have been classified as at increased risk. After 2397 and 492 vCJD-free years of follow-up, respectively, no deaths in either at-risk cohort were of vCJD-related causes. CONCLUSIONS: The at-risk cohorts have survived disease-free far longer than the estimated incubation time for dietary-acquired vCJD (donors) and transfusion-acquired disease (other recipients). However, due to our still limited understanding of, and a lack of a reliable test for, asymptomatic vCJD infection, public health follow-up is necessary for continued monitoring of at-risk cohorts.

Variant CJD and blood transfusion: are there additional cases?

BACKGROUND AND OBJECTIVES: In this study, we compare variant Creutzfeldt-Jakob disease (vCJD) cases definitely linked to blood transfusion, those with a history of blood transfusion in which no donor has developed vCJD and primary cases with no history of blood transfusion. The aim is to determine whether there are any differences in the demographics or clinical phenotype in these groups that might suggest additional cases of transfusion transmission of vCJD. MATERIALS AND METHODS: All cases of vCJD who are old enough to donate blood (i.e. >17 years old) are notified to the UKBTS at diagnosis, regardless of whether they are known to have a blood donation history. A search is then made for donor records and, if found, all components produced and issued to hospitals are identified and their fate determined. Recipient details are then checked against the NCJDRSU register to establish whether there is a match between these individuals and patients who have been diagnosed with vCJD. In the reverse study, attempts are made to trace the donors to all cases reported to have received a blood transfusion and donors' details are checked against the register to determine if any have developed vCJD. RESULTS: Of the 177 cases of vCJD diagnosed in the UK as of 1 February 2014, the TMER study identified 15 cases reported to have received a blood transfusion. Transfusion records were unavailable for 4 of these cases, all pre-1980, and in one other case there was no transfusion recorded in the medical notes. Transfusion records were found for 10 cases. One case transfused at symptom onset was excluded from this analysis. The mean age at onset of symptoms of the remaining nine transfusion recipients (four female and five male) was 42·9 years; 57·6 years in the three known transfusion-transmitted cases and 35·5 years in the six not linked cases. In one of these cases, details of components transfused were unavailable, and the remaining five cases received a total of 116 donor exposures with 112 donors identified, none of whom is known to have developed clinical vCJD. To date, five of the 112 identified donors have died and none was certified as dying of vCJD or any other neurological disorder. Two of the transfusion-transmitted cases did not fulfil diagnostic criteria for probable vCJD during life but were confirmed at post-mortem. Both cases were in the older age range (68 and 74 years, respectively), and neither had a positive MRI brain scan. The remaining cases all fulfilled the criteria for the diagnosis of vCJD in life, but two of these had atypical features and were older than the expected age at onset for vCJD. CONCLUSION: In conclusion, it is possible that one or more of the vCJD cases that received a blood transfusion derived from an individual not known to have vCJD were infected by the blood transfusion. However, the evidence for this is weak, and the absence of a past history of transfusion in most cases of vCJD excludes a large number of unrecognised transfusion-transmitted cases.

A conserved peptide in West Nile virus NS4A protein contributes to proteolytic processing and is essential for replication.

The West Nile virus strain Kunjin virus (WNV(KUN)) NS4A protein is a multifunctional protein involved in membrane proliferation, stimulation of cellular pathways, and evasion of host defense and is a major component of the WNV(KUN) RNA replication complex. We identified a highly conserved region ((120)P-E-P-E(123)) upstream of the viral protease dibasic cleavage site and investigated whether this motif was required for WNV(KUN) replication. Single point mutations to alanine and a PEPE deletion mutation were created in a full-length infectious WNV(KUN) molecular clone. All mutations drastically impaired viral replication and virion production, except that of the P122A mutant, which was slightly attenuated. These mutations were subsequently transferred to a WNV(KUN) replicon to specifically assess effects on RNA replication alone. Again, all mutants, except P122A, showed severely reduced negative-sense RNA production as well as decreased viral protein production. Correspondingly, immunofluorescence analyses showed a lack of double-stranded RNA (dsRNA) labeling and a dispersed localization of the WNV(KUN) proteins, suggesting that replication complex formation was additionally impaired. Attempts to rescue replication via conservative mutants largely failed except for substitution of Asp at E121, suggesting that a negative charge at this residue is equally important. Analysis of viral protein processing suggested that cleavage of the 2K peptide from NS4A did not occur with the mutant constructs. These observations imply that the combined effects of proline and negatively charged residues within the PEPE peptide are essential to promote the cleavage of 2K from NS4A, which is a prerequisite for efficient WNV replication.

The role of cerebrospinal fluid 14-3-3 and other proteins in the diagnosis of sporadic Creutzfeldt-Jakob disease in the UK: a 10-year review.

BACKGROUND: It is 10 years since the detection of cerebrospinal fluid (CSF) 14-3-3 was included in the diagnostic criteria for sporadic Creutzfeldt-Jakob disease (sCJD) by the WHO. Since that time, other CSF proteins, such as S100b and tau protein, have been proposed as surrogate markers for sCJD. The authors aimed to investigate the diagnostic value of each of these three proteins. METHODS: CSF samples collected from patients who were referred to the National CJD Surveillance Unit as suspected cases of sCJD during the period 1997-2007 were analysed for 14-3-3, S100b and tau protein. The sensitivity, specificity, positive predictive value and negative predictive value of each of these markers, either alone or in combination for the diagnosis of sCJD, were assessed. The impact of CSF 14-3-3 analysis on the case classification of sCJD was investigated. RESULTS AND DISCUSSION: CSF 14-3-3 had the greatest sensitivity (86%) when compared with tau protein (81%) and S100b (65%). The combination of a positive CSF 14-3-3 or an elevated tau protein with a raised S100b had the highest positive predictive power for sCJD. During the study period, 100 patients were classified as probable sCJD solely on the basis of the clinical features and a positive CSF 14-3-3. The most sensitive marker for sCJD was a positive CSF 14-3-3. The analysis of CSF 14-3-3 plays a crucial role in the case classification of sCJD.

Variant Creutzfeldt-Jakob disease and exposure to fractionated plasma products.

BACKGROUND: The risk to public health of onward transmission of variant Creutzfeldt-Jakob disease (vCJD) via blood transfusion and plasma product administration is of on-going concern, particularly with the recent reported detection of abnormal prion protein in a person with haemophilia. OBJECTIVES: To describe the history of fractionated plasma product exposure in clinical cases of vCJD in the UK. METHODS: Through examination of records held at the National CJD Surveillance Unit (from relatives, general practices and hospitals). RESULTS: Nine out of 168 UK vCJD cases had a history of receipt of fractionated plasma products on 12 different occasions (1 pre-vCJD risk in 1970, the remaining between 1989-1998). According to the UK CJD Incident Panel risk assessment criteria, 11 were low-risk products and one was low or medium risk. CONCLUSION: It is unlikely that any of the UK vCJD clinical cases to date were infected through exposure to fractionated plasma products. However, the possibility that such transmission may result in vCJD cases in the future cannot be excluded.

The Baby Friendly Hospital Initiative and breast feeding rates in Scotland.

OBJECTIVE: To examine the effect of the Baby Friendly Hospital Initiative on breast feeding rates in Scotland. DESIGN: Observational study using an annual survey of progress towards the WHO/UNICEF Baby Friendly Hospital Initiative and routinely collected breast feeding rates gathered on the Guthrie Inborn Errors Screening card at 7 days of postnatal age. SETTING: Scotland, UK, population 5.1 million, with about 53 000 births a year. PARTICIPANTS: All 33 maternity units with over 50 births per annum and 464,246 infants born in Scotland between 1995 and 2002. MAIN OUTCOME MEASURES: Baby Friendly status of each maternity unit at the time of an infant's birth: certificate of commitment, UK standard award, and breast feeding at 7 days postnatal age. RESULTS: Babies born in a hospital with the UK Baby Friendly Hospital Initiative standard award were 28% (p<0.001) more likely to be exclusively breast fed at 7 days of postnatal age than those born in other maternity units after adjustment for mother's age, deprivation, hospital size, and year of birth. From 1995, breast feeding rates had increased significantly faster in hospitals with Baby Friendly status by 2002: 11.39% (95% confidence interval 10.35 to 12.43) v 7.97% (95% confidence interval 7.21 to 8.73). CONCLUSION: Being born in a hospital that held the award increased the chance of being breast fed. All maternity units should be encouraged to undertake the significant strategic and practical changes required to achieve UK Baby Friendly Hospital Initiative standard status.

Conserved amino acids within the N-terminus of the West Nile virus NS4A protein contribute to virus replication, protein stability and membrane proliferation.

The West Nile virus strain Kunjin virus (WNVKUN) NS4A protein is a multifunctional protein involved in many aspects of the virus life-cycle and is a major component of the WNVKUN replication complex (RC). Previously we identified a conserved region in the C-terminus of NS4A regulating proteolytic processing and RC assembly, and now investigate key conserved residues in the N-terminus of NS4A and their contribution to WNVKUN replication. Mutation of P13 completely ablated replication, whereas, mutation of P48 and D49, near the first transmembrane helix, and G66 within the helix, showed variable defects in replication, virion secretion and membrane proliferation. Intriguingly, the P48 and G66 NS4A mutants resulted in specific proteasome depletion of NS4A that could in part be rescued with a proteasome inhibitor. Our results suggest that the N-terminus of NS4A contributes to correct folding and stability, essential for facilitating the essential roles of NS4A during replication.

X-linked and FSH dystrophies in one family.

A family is reported in which the father was affected by facioscapulohumeral muscular dystrophy FSHD. One son was affected by Duchenne muscular dystrophy (DMD). The second son died at the age of 3 yr of a severe primary muscle disease and it is suggested that this was the outcome of dual expression of the two conditions.

Malignant meningitis: a rational approach to cerebrospinal fluid cytology.

AIM: To clarify laboratory guidelines for cerebrospinal fluid (CSF) cytology. METHODS: Clinical and pathological data relating to 54 patients with cytologically malignant cells in the CSF were reviewed, together with CSF cell counts and protein measurements for 29 patients. Utilising this data, criteria were established for CSF cytology and validated by review of 100 patients in whom CSF cytology had not been carried out on the basis of these criteria. RESULTS: There was only one false positive diagnosis of malignancy on the basis of CSF cytology. All patients with malignant cells in the CSF fulfilled at least one of the following criteria: clinically known or suspected malignancy; raised cell count; raised protein concentration. In none of the 100 patients, in whom cytology was not performed, was the diagnosis of malignant meningitis missed. CONCLUSION: Cytology should be performed on CSF specimens from all patients with known, or suspected, malignancy, but in other cases, only if the cell count or protein concentration, or both, is raised.

Bleb formation and F-actin distribution during mitosis and tumor necrosis factor-induced apoptosis.

The murine cell line C3HA has been used extensively in studies of the cytopathology that accompanies TNF-induced cytolysis. This cell line undergoes an apoptic form of cell death characterized by plasma membrane blebbing and cytoplasmic boiling. Since plasma membrane blebs also appear on C3HA cells during mitosis, in this report we have compared these blebs with those that appear during apoptosis to determine whether they represent related structures. Our results reveal several differences. During mitosis, the blebs that appear are smaller and more heterogeneous in size than are those that appear during apoptosis. In addition, during mitosis bleb formation is preceded by the appearance of microvilli on the cell surface. No microvilli are observed during apoptosis. The staining pattern with rhodamine phalloidin also differed between mitotic and apoptic blebs, indicating a difference in their content of f-actin. The blebs that form during mitosis stained in a bright, uniform manner, suggesting an association with f-actin. In contrast, apoptic blebs were stained only at their base, the bleb itself being devoid of f-actin-associated staining. This difference may help explain why mitotic blebs are reintegrated into the cell surface, while the blebs that form during apoptosis are not.

Improved membrane preservation of flavivirus-infected cells with cryosectioning.

Ultra-cryomicrotomy and electron microscopy were used to investigate membranous structures in dengue virus-infected mammalian and insect cells. The cryo-sectioned samples displayed ultrastructure comparable to their resin-embedded counterparts with all previously identified virus-induced structures being observed. Structures not previously identified were also found. In particular, membrane-bound packets of vesicles, 100-200 nm in diameter were seen distributed throughout areas of virus-induced membrane proliferation. These packets were clearly distinct from virion arrays. Small smooth membrane vesicles, previously found to contain thread-like enclosures (M.L. Ng, J. Gen. Virol. 68 (1987) 577-582), were frequently observed to contain dense staining material, however the exact nature of this material remains unclear. Virus-induced modification of golgi-like and/or ER membranes was also observed and may represent early events in the generation of the smooth membrane vesicles seen during infection. We suggest that cryosectioning is the method of choice to investigate membrane rearrangement induced by this family of viruses and that a diamond knife and modified staining techniques, as utilised in this report, be employed to enhance morphology and section preservation.

Effects of body mass and genotype on avian degenerative joint disease pathology and articular cartilage proteoglycan distribution.

OBJECTIVE: The objective of this study was to investigate the role of body mass and genotype in the development of avian degenerative joint disease (DJD). METHODS: Layer strain and broiler strain fowl, fed either ad libitum or on a restricted diet, were kept under identical conditions for up to a year. At various time points cartilage samples were taken from the distal tibiotarsus (DTT), proximal tarsometatarsus, antitrochanter and proximal humerus. All samples were assessed for gross morphology and histopathology, and in some samples the cartilage proteoglycan distribution was investigated by Safranin O staining. RESULTS: Layer strain fowl did not develop DJD. Heavy ad libitum fed broiler strain fowl developed DJD earlier and more severely than lighter, feed restricted, broiler strain fowl. The articular surface of the DTT was worst affected by DJD. Safranin O staining of DTT samples (age 180 days) from the ad libitum fed broilers revealed variable proteoglycan distribution in the articular cartilage. Some areas were intensely stained throughout all zones, whereas other areas showed no staining in any zone. Age matched, non-diseased DTT samples from feed restricted broilers showed a more consistent staining pattern with little staining in the surface zone and more in the middle and deep zones. CONCLUSIONS: These data indicate that avian DJD is body mass mediated in broiler strain fowl, and that proteoglycan distribution is altered in diseased cartilage.

Changes in proteoglycan content of articular cartilage during avian degenerative joint disease.

OBJECTIVE: To determine the biochemical changes in articular cartilage composition associated with the development of avian degenerative joint disease (DJD) in ad libitum fed broiler fowl, in comparison to feed-restricted broilers and J-lin fowl (non-susceptible to DJD). METHODS: Articular cartilage from the distal tibiotarsus (DTT) was characterised up to age 180 days. Proteoglycan content was determined by uronic acid and sulphated glycosaminoglycan analysis, cellularity by assay for DNA content, and collagen content and crosslinking by hydroxyproline and pyridinoline analysis, respectively. RESULTS: Disease development was accompanied by increased hydration and proteoglycan content (particularly sulphated proteoglycans) and decreased cellularity, with no significant differences in either total collagen content or in mature collagen cross-linking. CONCLUSION: The biochemical features of avian DJD are similar to those observed in other animal models. This bipedal model is exceptional however since cartilage alterations occur spontaneously and in a load-dependent manner.

A leiomyosarcoma of the oesophagus presenting incidentally without dysphagia.

Leiomyosarcoma is a rare tumour that accounts for 0.5% oesophageal sarcomas. The most common presenting symptom is dysphagia. This report presents a case of oesophageal leiomyosarcoma in a 56-year-old Caucasian man found incidentally while being investigated for refractory cough. There was no history of dysphagia in spite of tumour mass occupying most of the oesophageal lumen. The leiomyosarcoma was managed successfully with surgical resection and adjuvant radiotherapy. The patient remains disease free after 15 months after surgical intervention. The unusual case presentation is discussed and the surgical management of this rare condition reviewed.

Birefringent periodicities in amphibian rod outer segments.

Birefringence variations, seen as regularly spaced altering light and dark rings (bands), have been observed along the length of unfixed, freshly isolated rod outer segments (ROS) of both Rana pipiens and Xenopus laevis. In our hands, the spatial frequency of the banding pattern is from 1.3-1.6u/band in Rana pipiens and 1.8-2.2u/band in Xenopus laevis ROS, both corresponding closely to determinations we made in the same animals of the quantity of new ROS disks added each day. To further probe this correlation, Xenopus laevis were maintained at 16 degrees C to lower the disk renewal rate. A similar correspondence was found, with the banding pattern and renewal rate both at 0.8-1.Ou/day. Further experiments involving Xenopus laevis placed on altered lighting cycles have suggested the existence of two normally superimposed periodicities. The more intense component is driven by the environmental light cycle, and thus may be regarded as diurnal. The less intense component is seen infrequently, suggesting lability, and apparently follows a 24-hour period in both constant light and darkness.

The anatomy of aneurysm-bearing circles of Willis.

The circle of Willis is known to exhibit considerable anatomical variability. The incidence of variations of the circle is said to be greater when an aneurysm is present, particularly one of the anterior communicating artery. In this study 40 circles of Willis bearing 51 saccular aneurysms were dissected and examined macroscopically. Forty circles from patients with other neurological diseases served as controls. Ninety-seven per cent of circles bearing aneurysms showed variations from a hypothetical "normal" structure, as did 85% of controls. The aneurysm group as a whole, and circles bearing anterior communicating aneurysms, showed a higher incidence of anterior cerebral artery asymmetry than controls (p less than 0.001), as did the latter when compared with circles bearing aneurysms at other sites (p less than 0.01), but there were no other significant differences.

Degenerative joint disease in poultry--differences in composition and morphology of articular cartilage are associated with strain susceptibility.

The morphology and basic biochemical composition of articular cartilage from two strains of fowl were examined. Broiler breeder fowl are considered susceptible to degenerative joint disease (DJD); histological examination of one-year-old broiler breeders showed in some samples, articular cartilage thinning, fibrillation and chondrocyte cluster formation, features considered typical of DJD. Examination of similar samples from laying strain fowl showed only minor age-related changes such as some slight cartilage thinning and very mild fibrillation. The articular cartilage from the broiler breeder birds was significantly more hydrated with a higher uronic acid content than that of the laying strain birds. In addition, unloaded articular surfaces such as the proximal humerus had significantly higher amounts of uronic acid than the loaded cartilage surfaces of the proximal tarsometatarsus and the distal tibiotarsus; this suggested that the joint loading may have a role in any biochemical differences found between joints and between strains of fowl. These findings concur with other reports in mammals that showed increased hydration and uronic acid in association with early DJD and in models of osteoarthritis (OA). Thus, despite some differences between avian and mammalian articular cartilage, studies on avian DJD may give insights into mammalian disease.