Multidimensional Analysis of the Adult Human Heart in Health and Disease using Hierarchical Phase-Contrast Tomography (HiP-CT).

Cardiovascular diseases (CVDs) are a leading cause of death worldwide. Current clinical imaging modalities provide resolution adequate for diagnosis but are unable to provide detail of structural changes in the heart, across length-scales, necessary for understanding underlying pathophysiology of disease. Hierarchical Phase-Contrast Tomography (HiP-CT), using new (4th) generation synchrotron sources, potentially overcomes this limitation, allowing micron resolution imaging of intact adult organs with unprecedented detail. In this proof of principle study (n=2), we show the utility of HiP-CT to image whole adult human hearts ex-vivo: one 'control' without known cardiac disease and one with multiple known cardiopulmonary pathologies. The resulting multiscale imaging was able to demonstrate exemplars of anatomy in each cardiac segment along with novel findings in the cardiac conduction system, from gross (20 um/voxel) to cellular scale (2.2 um/voxel), non-destructively, thereby bridging the gap between macroscopic and microscopic investigations. We propose that the technique represents a significant step in virtual autopsy methods for studying structural heart disease, facilitating research into abnormalities across scales and age-groups. It opens up possibilities for understanding and treating disease; and provides a cardiac 'blueprint' with potential for in-silico simulation, device design, virtual surgical training, and bioengineered heart in the future.

Tumor suppressor p53: analysis of wild-type and mutant p53 complexes.

It has been suggested that the dominant effect of mutant p53 on tumor progression may reflect the mutant protein binding to wild-type p53, with inactivation of suppressor function. To date, evidence for wild-type/mutant p53 complexes involves p53 from different species. To investigate wild-type/mutant p53 complexes in relation to natural tumor progression, we sought to identify intraspecific complexes, using murine p53. The mutant phenotype p53-246(0) was used because this phenotype is immunologically distinct from wild-type p53-246+ and thus permits immunological analysis for wild-type/mutant p53 complexes. The p53 proteins were derived from genetically defined p53 cDNAs expressed in vitro and also from phenotypic variants of p53 expressed in vivo. We found that the mutant p53 phenotype was able to form a complex with the wild type when the two p53 variants were cotranslated. When mixed in their native states (after translation), the wild-type and mutant p53 proteins did not exhibit any binding affinity for each other in vitro. Under identical conditions, complexes of wild-type human and murine p53 proteins were formed. For murine p53, both the wild-type and mutant p53 proteins formed high-molecular-weight complexes when translated in vitro. This oligomerization appeared to involve the carboxyl terminus, since truncated p53 (amino acids 1 to 343) did not form complexes. We suggest that the ability of the mutant p53 phenotype to complex with wild type during cotranslation may contribute to the transforming function of activated mutants of p53 in vivo.

Assessment of fitness for purpose of an insect bioassay using the desert locust (Schistocerca gregaria L.) for the detection of paralytic shellfish toxins in shellfish flesh.

We have developed a bioassay using 5th instar desert locusts (Schistocerca gregaria L.) for the detection of saxitoxin-the paralytic shellfish poison in shellfish flesh. The bioassay procedure is to inject 10 locusts with a shellfish extract, and assess their conditions at time points up to 2h post injection, looking for an endpoint of paralysis. From the proportion responding, the equivalent dose of pure saxitoxin could be estimated. Performance characteristics of the bioassay were assessed using shellfish samples spiked with saxitoxin, and we found the bioassay could detect and quantify toxin levels in the range of regulatory relevance. Relative toxicities of selected saxitoxin analogues differed from those reported in mammalian systems. Variation for repeatability conditions was acceptable but variation was higher under reproducibility conditions. This was related to (a) batches of insects from different suppliers, (b) different operators, and (c) different observers assessing the endpoint. We also noted adverse reactions with some shellfish species. These problems may be resolved by further refinement of the method and operator training, before formal validation. Nevertheless, we suggest the method potentially offers a simple, ethically acceptable, broad-specificity functional bioassay, which is desirable in any toxin-monitoring programme.

Atrioventricular septal defects diagnosed in fetal life: associated cardiac and extra-cardiac abnormalities and outcome.

OBJECTIVES: We sought to establish the outlook for fetuses diagnosed with atrioventricular septal defect (AVSD) prenatally and its relation to additional cardiac, extracardiac and chromosomal abnormalities. BACKGROUND: Prediction of likely outcome of AVSD presenting prenatally is complicated by the wide variation in associated features. METHODS: Computerized records from 14,726 pregnancies referred to a fetal cardiology center were reviewed retrospectively. Pathological reports, postnatal records, follow-up inquiries and review of echocardiographic video recordings supplemented analysis of the records for all those with AVSD. RESULTS: Atrioventricular septal defect was confirmed in 301 fetuses. Eighty-six (39%) of the 218 with known karyotype had trisomy 21, and 21/218 (10%) had other chromosome abnormalities. Right isomerism occurred in 37/301 (12%) fetuses, left isomerism in 62 (20%), mirror image atrial arrangement in 2 (1%), and 200 (67%) had usual arrangement. Atrioventricular septal defect occurred without any other intracardiac abnormality in 155 fetuses (51%). Extracardiac abnormalities and nonkaryotypic syndromes were evident in 40 fetuses (13%, confidence interval [CI] 9.5-17.1%). Uncomplicated cardiac anatomy was significantly associated with the presence of karyotype abnormality (p < 0.0001). Parents opted for termination of pregnancy in 175/298 (58.5%). For the continuing pregnancies, Kaplan-Meier estimates for live birth, survival past the neonatal period and survival to three years were 82% (CI 75.3-88.9%), 55% (CI 46.0%-0/64.3%) and 38% (CI 27.1-48.6%), respectively. Fetal hydrops and earlier year of diagnosis were independent variables with adverse influence on survival. CONCLUSIONS: Despite some improvements in the outlook for AVSD diagnosed prenatally, the overall prognosis remains considerably poorer than that implied from surgical series. The detection of associated cardiac and extracardiac abnormalities is important in order to give the best indication of the likely outcome when counseling parents.

Prospective diagnosis of 1,006 consecutive cases of congenital heart disease in the fetus.

OBJECTIVE: This report describes our experience with fetal congenital heart disease since 1980. BACKGROUND: Knowledge and expertise in the diagnosis, management and natural history of fetal congenital heart disease is increasingly demanded by both obstetricians and parents. The analysis of a large series should help the pediatric cardiologist to provide this service. METHODS: The notes of 1,006 patients, where a prospective diagnosis of fetal congenital heart disease was made, were reviewed. The reason for referral, the diagnosis made, the accuracy of diagnosis, the fetal karyotype and the outcome of the pregnancy were noted. The cases were grouped into malformation categories, and the spectrum of disease seen was compared with that found in infants. RESULTS: Most fetal cardiac anomalies are now suspected by the ultrasonographer during obstetric scanning. A different incidence of abnormalities is seen compared with that expected in infants. Chromosomal anomalies were more frequent in the fetus than in live births. The accuracy of diagnosis was good. The survival rate after diagnosis was poor because of frequent parental choice to interrupt pregnancy and the complexity of disease. CONCLUSIONS: A large experience with fetal congenital heart disease allows the spectrum of disease to be described with accuracy and compared with that in infancy. Knowledge of the natural history of heart malformations when they present in the fetus allows accurate counseling to be offered to the parents. If the trend in parental decisions found in this series continues, a smaller number of infants and children with complex cardiac lesions will present in postnatal life.

Isolated congenitally complete heart block attributable to combined nodoventricular and intraventricular discontinuity.

Intraventricular together with atrial-axis and nodoventricular discontinuity, in which various parts of the conduction system are replaced by fibrous or fatty tissue, constitute the three major pathological categories of isolated congenitally complete heart block. Intraventricular discontinuity is distinctly rare, with only two previous cases reported in the literature, one of which was associated with a familial history of heart block. The cardiac conduction systems of two cases of isolated congenitally complete heart block were serially sectioned and analyzed histopathologically. The findings were correlated with the clinical features, in particular, the family histories and maternal serum anti-Ro antibodies. Both cases, a 9-day-old neonate and an 8-year-old schoolgirl, showed a combination of nodoventricular and intraventricular discontinuity, with absence of the atrioventricular penetrating bundle, the entire right, and the proximal portion of the left bundle branch. The branching bundle was absent in the first case and replaced by fatty tissue in the second. In contrast to the commoner atrial-axis discontinuity in which the atrioventricular node itself is usually replaced by fibrous or fatty tissue with variable involvement distally, the sinus node, and in particular, the atrioventricular node were normal in both of our cases. There was no family history in either case, whereas tests for the maternal serum anti-Ro antibody were positive in the first but negative in the second case. Intraventricular discontinuity as a cause of isolated congenitally complete heart block is very rare. In our cases, it co-existed with nodoventricular discontinuity. It can be sporadic, familial, or associated with positive maternal serum anti-Ro antibodies.

Clusters within a general adult population of alcohol abstainers.

BACKGROUND: Our previous study found that alcohol abstainers use acute services more and preventative services less than safe level drinkers. The observed relationships between four categories of alcohol consumption and service use were J-shaped for acute services and inverted J-shaped for preventive services. The aim of this paper was to further investigate these relationships. METHODS: The design was a health and lifestyle survey of 41 000 randomly-sampled adults in SE England. The response rate was 60%. Distinctive subgroups within the alcohol abstainer group were investigated using cluster analysis, based on socio-demographic and health status variables. Odds ratios for services use for the abstainer clusters, and three alcohol consumption groups were estimated from a logistic regression model which included age, social class, ethnic group, employment status, household composition, whether the respondent was a carer, smoking habit, use of private health insurance, and health status. RESULTS: Two clusters were formed for both males and females. Cluster 1 comprised, on average, older, frailer, and more disabled people. Cluster 2 comprised younger, healthier people, a greater proportion of whom belonged to ethnic minority groups. Cluster 2 had similar rates of use of Accident & Emergency, GP, optician, and dental services compared with safe level drinkers. Cluster 1's rates differed from those of both Cluster 2 and safe level drinkers in almost all instances. CONCLUSIONS: The J- and inverted J-shaped relationships between alcohol consumption and service use are partly explained by a subgroup of abstainers who are older, of less good health, and who use hospital, clinic, and domiciliary healthcare services much more than safe level drinkers.

Histological changes in the left and right ventricle in hearts with Ebstein's malformation and tricuspid valvar dysplasia: A morphometric study of patients dying in the fetal and perinatal periods.

Both Ebstein's malformation and the related tricuspid valvar dysplasia are often associated with tricuspid regurgitation, and impaired right ventricular function may develop. Impaired function of the left ventricle in Ebstein's malformation has also been described. Interstitial fibrosis has been shown in the right and left ventricles of hearts with Ebstein's malformation from neonates, children, and adults. The objective of this study was to determine whether interstitial fibrosis seen in Ebstein's malformation is an intrinsic part of the congenital malformation or is acquired. From the fetal and perinatal periods, we compared 13 hearts with Ebstein's malformation (6 isolated and 7 with additional abnormalities) and 11 with tricuspid valvar dysplasia (3 isolated and 8 with additional abnormalities) with 16 controls. Three adult cases of isolated Ebstein's malformation in patients aged 17 to 20 years, were compared with 5 controls. The percentage of interstitial fibrous tissue and the thickness of the endocardium in the right and left ventricles were measured using histomorphometry. There were similar findings in Ebstein's malformation and tricuspid valvar dysplasia. Of 24 fetal and perinatal cases, 23 had normal interstitial fibrous tissue. Interstitial fibrosis was found in the right ventricle of only 1 perinatal heart with Ebstein's malformation and pulmonary stenosis. Of the 9 fetal cases, 4 had minimal right ventricular endocardial thickening (up to 10 µm). The left ventricular endocardium was normal in this group. Of the 6 perinatal cases with isolated Ebstein's malformation or tricuspid valvar dysplasia, 4 had right and 2 had left ventricular endocardial thickening (up to 345 µm). Of the 3 adult hearts with Ebstein's malformation, 2 had right ventricular endocardial thickening (47 and 225 µm) and 2 had right and 1 had left ventricular interstitial fibrosis. These results indicate that in both Ebstein's malformation and tricuspid valvar dysplasia the endocardial thickening develops in perinatal life, and in Ebstein's malformation the interstitial fibrosis develops in later life.

Surgical anatomy of aorto-left ventricular tunnel.

BACKGROUND: Owing to the rarity of aorto-left ventricular tunnel, surgical experience with this condition is generally limited. The anatomic configuration remains to be clarified in the light of better understanding of the normal aortic root. METHODS: Two autopsied hearts with aorto-left ventricular tunnel were examined and compared with four normal heart specimens. The normal hearts were sectioned in a variety of planes to display the ventriculoarterial junction. RESULTS: The leaflets of the pulmonary valve in both normal and abnormal hearts have semilunar attachments to a sleeve of freestanding ventricular musculature, the infundibulum. An extensive fibrofatty tissue plane then interposes between the freestanding infundibulum and the aortic sinuses. The aorto-left ventricular tunnels in the abnormal hearts pass within this tissue plane. The aortic orifice of the tunnel is distal to the level of the sinutubular junction, whereas the ventricular orifice is located within the interleaflet triangle between the right and left aortic sinuses. CONCLUSIONS: Aorto-left ventricular tunnels bypass the normal ventriculoarterial junction but do not penetrate the septal musculature. This has implications for the fine-tuning of surgical repair.

The use of acute and preventative medical services by a general population: relationship to alcohol consumption.

AIMS: To investigate the hypothesis that increasing alcohol consumption is accompanied by increasing use of acute, but decreasing use of preventative, medical services among the general population. DESIGN AND PARTICIPANTS: Health and life-style survey of 41,000 randomly-sampled adults in SE England who self-completed a validated questionnaire covering socio-demographics, alcohol and tobacco usage and use of acute (A&E department and general practitioner) and preventative (dental, optician, mammography and cervical cytology) services: the response rate was 60%. MEASUREMENTS: Comparative use of acute and preventative health care services by patients with varying consumption of alcoholic beverages. This was estimated by the odds ratio for service use, after correcting for the following confounding variables; age, social class, ethnic group, employment status, whether lives with children or with other adults, whether is a career, limiting long-term illness, depression status, smoking habit and use of private health insurance. FINDINGS: There was increased use of accident and emergency services by the harmful and intermediate drinking groups compared with the safe drinking group. Male abstainers attended their A&E departments more frequently than 'safe limit' drinkers. With respect to preventative services, both male and female abstainers and harmful drinkers used dental services less than safe limit drinkers. For females, mammography and cervical cytology services were less frequently used by abstainers and by harmful drinkers. CONCLUSIONS: This study supports the generally held view that heavy alcohol consumers are disproportionate users of acute medical services but they are relative under-users of preventative medical care services. Alcohol abstainers are also over-users of acute services, but under-users of preventative services. These latter observations are relevant to the claims that moderate alcohol consumers have lower apparent morbidity and mortality rates compared to abstainers.

Decrease in timed balance test scores with aging.

This study investigated whether 184 volunteers from 20 to 79 years of age could perform eight timed balance tests and examined the relationship between test performance and age. All subjects were able to balance with their feet together and eyes closed for 30 seconds. The ability to balance on the right and left legs did not differ significantly. Subjects over 60 years of age were unable to balance on one leg, particularly when their eyes were closed, for as long a period as younger subjects. The Pearson product-moment and Spearman correlations of age and duration of one-legged balance were -.65 and -.71 (eyes opened) and -.79 and -.75 (eyes closed). The findings suggest that when timed balance tests are performed as a part of a patient's neurologic examination, the results should be interpreted in light of the patient's age. Information is provided to assist in this interpretation.

Early mobilization following carpal tunnel release. A prospective randomized study.

A prospective randomized study was undertaken of 50 consecutive patients undergoing surgery for idiopathic carpal tunnel syndrome to determine the value of splintage of the wrist following open carpal tunnel release. Patients were randomized to either be splinted for 2 weeks following surgery or to begin range-of-motion exercises on the first post-operative day. Subjects were evaluated at 2 weeks, 1 month, 3 months, and 6 months after surgery by motor and sensory testing, physical examination, and a questionnaire. Variables assessed included date of return to activities of daily living, dates of return to work at light duty and at full duty, pain level, grip strength, key pinch strength, and occurrence of complications. Patients who were splinted had significant delays in return to activities of daily living, return to work at light and full duty, and in recovery of grip and key pinch strength. Patients with splinted wrists experienced increased pain and scar tenderness in the first month after surgery; otherwise there was no difference between the groups in the incidence of complications. We conclude that splinting the wrist following open release of the flexor retinaculum is largely detrimental, although it may have a role in preventing the rare but significant complications of bowstringing of the tendons or entrapment of the median nerve in scar tissue. We recommend a home physiotherapy programme in which the wrist and fingers are exercised separately to avoid simultaneous finger and wrist flexion, which is the position most prone to cause bowstringing.

The spectrum of fetal cardiac malformations.

Increasingly, paediatric cardiologists are called upon to diagnose cardiac malformations prenatally. In the main, the types of malformation seen during fetal life will be similar to those documented postnatally, but the frequency with which they are encountered, as well as the views that can be used for diagnosis, will be different. This review aims to describe the anatomic spectrum of malformations seen in 917 fetal hearts examined consecutively following prenatal diagnosis. The distribution of anomalies is illustrated in terms of a simple sweep through the fetal thorax passing from the four-chamber plane to the outflow tracts, and then to more cranial views of the mediastinum. Two-thirds of the anomalies described would have been detectable in the four-chamber plane. Some, such as tricuspid valvar abnormalities, will alter the normal appearances of the four-chambers dramatically. In terms of the overall spectrum, however, such obvious abnormalities only form a minor part of the total number. Others, such as atrioventricular septal defect, will often require closer inspection of intracardiac anatomy, but will make up a large proportion of the entire cohort. Up to one third of the anomalies in the series would have required views more cranial to the four-chamber plane of section. In these, it would have been necessary to examine the nature of the left ventricular outflow tract, the crossing of the two outflows, or else the arterial arches in order to secure detection. In the fetus, these and other planes must be considered by the echocardiographer in order completely to detect and document the entire spectrum of cardiac abnormalities likely to be encountered.

Use of casts in the necropsy diagnosis of fetal congenital heart disease.

OBJECTIVE: To evaluate a casting technique in the interpretation of fetal cardiac anatomy. DESIGN: In 32 fetuses, the echocardiographic and cast features were compared and correlated. PATIENTS: Three normal fetal heart specimens from spontaneous abortuses and 32 specimens from spontaneous or induced abortions with congenital heart malformation. RESULTS: There was close correlation between the echocardiographic and anatomical features in 32 abnormal fetuses studied. In some, additional features of diagnosis could be displayed on the cast and the relative sizes of the cardiac structures could be appreciated and defined. CONCLUSIONS: With increasing echocardiographic detection of congenital heart disease in early prenatal life, an increasing number of fetal heart specimens of small size are dissected for pathological confirmation. The use of silicone rubber casts to reproduce the internal anatomy proved a useful addition to dissection, providing a three dimensional model of the cardiac defect.

Atresia or absence of the left-sided atrioventricular connection in the fetus: echocardiographic diagnosis and outcome.

Our objective was to describe the appearance of atresia of the left-sided atrioventricular valve on the four-chamber view of the fetal heart. Twenty-four cases of atresia of the left-sided atrioventricular connection were detected over a 2-year period. Cases were analyzed according to the appearance of the four-chamber view and the connections of the great arteries. All cases were referred because the left-sided ventricle was smaller than the right. There were five different groups of cardiac anomaly detected. Associated findings varied according to the diagnostic group. It was concluded that it is possible to identify atresia of the left-sided atrioventricular connection accurately during obstetric ultrasound examination by evaluation of the four-chamber view. However, the appearance of this view varies with the diagnosis. Each diagnostic group has different implications in terms of associated extracardiac malformations and postnatal management.

Concordance for hypoplastic left heart syndrome in a monochorionic twin pregnancy.

The risk of structural heart disease is significantly higher in twin pregnancies than in singleton pregnancies, but the concordance rate has been found to be relatively low, even in monochorionic pregnancies. This is the first report of a monochorionic twin pregnancy concordant for hypoplastic left heart syndrome (HLHS), the diagnosis having been made by fetal echocardiography at 15 weeks' gestation. The findings were confirmed at necropsy at 17 weeks' gestation, following termination of pregnancy. Both twins had mitral and aortic atresia, with severely hypoplastic aortic arches. This report adds weight to there being a genetic component to the cause of HLHS in some cases and illustrates how the findings from early fetal echocardiography with postmortem follow up can help to extend the understanding of the aetiology of this condition.

Cardiac tumours in intrauterine life.

Since 1980, 11 examples of cardiac tumour have been detected in the fetus out of a total of 794 congenital cardiac malformations. Patients were referred because of fetal hydrops in two, a family history of tuberous sclerosis in two, and because of the detection of a tumour mass during a scan at the local hospital in seven. The gestational age range at presentation was from 20-34 weeks. Of eight fetuses where death occurred, the histological type was rhabdomyoma in seven and teratoma in one. In seven cases, the lesion appeared single and in four there were multiple tumours. In two of the cases of rhabdomyoma, other family members had evidence of tuberous sclerosis. Termination of pregnancy took place in four cases; of seven continuing pregnancies, spontaneous intrauterine death occurred in four, and three children are still alive. Two of the three survivors has the clinical picture of tuberous sclerosis. The last case is as yet only 1 month old. In summary, even where the lesion is single, the most likely diagnosis in fetal cardiac tumour is rhabdomyoma, with associated tuberous sclerosis. However, the characteristic features of this latter condition may not become evident until some months after birth.

Investigation of bias after data linkage of hospital admissions data to police road traffic crash reports.

RESEARCH QUESTION: Does a database of hospital admission data linked to police road traffic accident (RTA) reports produce less biased information for the injury prevention policymaker, planner, and practitioner than police RTA reports alone? DESIGN: Data linkage study. STUDY POPULATION: Non-fatal injury victims of road traffic crashes in southern England who were admitted to hospital. DATA SOURCES: Hospital admissions and police RTA reports. MAIN OUTCOME MEASURES: The estimated proportion of road traffic crashes admitted to hospital that were included on the linked database; distributions by age, sex, and road user groups: (A) for all RTA injury admissions and (B) for RTA serious injury admissions defined by length of stay or by nature of injury. RESULTS: An estimated 50% of RTA injury admissions were included on the linked database. When assessing bias, admissions data were regarded as the "gold standard". The distributions of casualties by age, sex, and type of road user showed major differences between the admissions data and the police RTA injury data of comparable severity. The linked data showed smaller differences when compared with admissions data. For RTA serious injury admissions, the distributions by age and sex were approximately the same for the linked data compared with admissions data, and there were small but statistically significant differences between the distributions across road user group for the linked data compared with hospital admissions. CONCLUSION: These results suggest that investigators could be misinformed if they base their analysis solely on police RTA data, and that information derived from the linked database is less biased than that from police RTA data alone. A national linked dataset of road traffic crash data should be produced from hospital admissions and police RTA data for use by policymakers, planners and practitioners.