Early axonal contacts during development of an identified dendrite in the brain of the zebrafish.

We have identified the initial synaptic contacts made onto the Mauthner (M) cell, an identified neuron that arises during early development of the zebrafish hindbrain. The contacts are made by a small bundle of pioneering trigeminal sensory axons onto the M cell soma before it forms dendrites. The sensory bundle is then partially enveloped by the M cell. The lateral dendrite appears at about the site of the contact, and eventually the trigeminal inputs are shifted to its trunk. As the dendrite elongates, other sensory contacts are made on its distal regions, sequentially from the acoustico-vestibular nerve and the lateral line nerves. To learn whether the earliest inputs induce the initial outgrowth of the M cell dendrite, we ablated the trigeminal neurons by laser irradiation before they contacted the M cell. Morphogenesis of the M cell, including its dendrite, appeared normal.

Collection and determinants of patient reported outcome measures in haemodialysis patients in Scotland.

BACKGROUND/INTRODUCTION: Patient reported outcome measures (PROMs) can evaluate the quality of health in patients with established renal failure. There is limited experience of their use within national renal registries. AIM: To describe the Scottish Renal Registry's (SRR) experience of collecting PROMS in the haemodialysis population and correlate PROMS to demographic and clinical parameters. DESIGN: Retrospective observational cross-sectional study. METHODS: Haemodialysis patients in Scotland were invited to complete the KDQOL™-36 questionnaire on the day of the annual SRR census in 2015 and 2016. Questionnaires were linked to census demographic and clinical variables. RESULTS: In 2016, 738 questionnaires were linked to census data (39% of prevalent haemodialysis population). Response rates differed with age (≥ 65 years 42%, < 65 years 36%) [χ2P = 0.006]; duration of renal replacement therapy (<1 year 46%, ≥1 < 5 years 38%, ≥ 5 years 33%) [χ2P = 0.002] and social class (Scottish Index of Multiple Deprivation (SIMD) Class 1 32%, Class 2 41%, Class 3 40%, Class 4 48%, Class 5 40%) [χ2P < 0.001]. There were significant differences in PROMs with age, SIMD quintile and primary renal diagnosis. Achieving a urea reduction ratio of >65% and dialysing through arteriovenous access were associated with significantly higher PROMs. PROMs were not affected by haemoglobin or phosphate concentration. DISCUSSION/CONCLUSIONS: Routine collection of PROMs is feasible and can identify potentially under-recognized and treatable determinants to quality of life. The association between attaining recommended standards of care and improved PROMs is striking. Individual and population-wide strategies are required to improve PROMs.

End-stage renal disease and survival in people with diabetes: a national database linkage study.

BACKGROUND: Increasing prevalence of diabetes worldwide is projected to lead to an increase in patients with end-stage renal disease (ESRD) requiring renal replacement therapy (RRT). AIM: To provide contemporary estimates of the prevalence of ESRD and requirement for RRT among people with diabetes in a nationwide study and to report associated survival. METHODS: Data were extracted and linked from three national databases: Scottish Renal Registry, Scottish Care Initiative-Diabetes Collaboration and National Records of Scotland death data. Survival analyses were modelled with Cox regression. RESULTS: Point prevalence of chronic kidney disease (CKD)5 in 2008 was 1.63% of 19 414 people with type 1 diabetes (T1DM) compared with 0.58% of 167 871 people with type 2 diabetes (T2DM) (odds ratio for DM type 0.97, P = 0.77, on adjustment for duration. Although 83% of those with T1DM and CKD5 and 61% of those with T2DM and CKD5 were receiving RRT, there was no difference when adjusted for age, sex and DM duration (odds ratio for DM type 0.83, P = 0.432). Diabetic nephropathy was the primary renal diagnosis in 91% of people with T1DM and 58% of people with T2DM on RRT. Median survival time from initiation of RRT was 3.84 years (95% CI 2.77, 4.62) in T1DM and 2.16 years (95% CI: 1.92, 2.38) in T2DM. CONCLUSION: Considerable numbers of patients with diabetes continue to progress to CKD5 and RRT. Almost half of all RRT cases in T2DM are considered to be due to conditions other than diabetic nephropathy. Median survival time for people with diabetes from initiation of RRT remains poor. These prevalence data are important for future resource planning.

Sensory neuron growth cones comigrate with posterior lateral line primordial cells in zebrafish.

The development of neuromasts and sensory neurons of the posterior lateral line was studied in zebrafish (Brachydanio rerio) in order to determine the relationship between growing axons of sensory neurons and the migratory cellular primordium of midbody line neuromasts. Scanning electron microscopy revealed that a primary system of six neuromasts develops during the second day after fertilization and evidence is presented that these arise from cells of a migratory primordium. The primordium is first detected in the postauditory region immediately adjacent to the developing sensory ganglion. Growth cones of posterior lateral line sensory neurons are found within the premigratory primordium when it is adjacent to the ganglion. At later times growth cones of these sensory neurons are found within the primordium as it migrates caudally along the midbody line. These results demonstrate that although the growth cones of the sensory neurons grow over a considerable distance to their final destination, they are never very far from their target cells (or target cell precursors), which migrate with them and may even lead them.

Segmental homologies among reticulospinal neurons in the hindbrain of the zebrafish larva.

We have examined the morphology of identified reticulospinal neurons in larval zebrafish by retrogradely labeling them with horseradish peroxidase. We described the morphology of 27 different types of reticulospinal neurons found in the hindbrain 5 days after fertilization. Nineteen of these types are present as single identified neurons on each side of the brain; the others are present as pairs or small groups of cells. The hindbrain reticulospinal neurons are present in seven bilateral clusters that are spaced periodically along the neuraxis. Each cluster contains two to five different types of reticulospinal neurons. Cells with similar morphological features are found in adjacent clusters. By considering cell position within the cluster and axon pathway, nearly all of the cells can be assigned to one of about seven serially repeated classes. Independent morphological features of the cells support the same classification. We propose that the clusters represent hindbrain segments and that the neurons of the same class that are present in the different clusters are segmental homologues. Assuming that this series evolved by successive duplications and divergence of the primitive segments, we have analyzed the changes that may have occurred during the evolution of each new segment. Changes between ipsilaterally and contralaterally projecting axons may have occurred several times during the evolution of the series. In addition, cells may have been added or deleted.

T reticular interneurons: a class of serially repeating cells in the zebrafish hindbrain.

We describe a class of reticular neurons, named T interneurons after the branching pattern of their axons, in young larvae of the zebrafish Brachydanio rerio. The cells were identified by filling them with HRP from application sites within the CNS. A serially repeating set of about ten such neurons is present in a longitudinal column on each side of the caudal hindbrain. The T axons project across the midline, and branches course both rostrally and caudally within the medial longitudinal fascicle (mlf). The cells receive synaptic input from the Mauthner neurons, from unidentified axons in the mlf, and perhaps from trigeminal sensory fibers. They project to cranial and pectoral motor nuclei. T interneurons appear to be homologous to giant fiber neurons in the hatchetfish and to some of the cranial relay neurons in the goldfish. We discuss a possible functional role and comparative implications of their distribution in the hindbrain.

Anatomy of the posterior lateral line system in young larvae of the zebrafish.

We studied the anatomy of neuromasts, afferent sensory neurons, and efferent neurons of the midbody branch of the posterior lateral line in larvae of the zebrafish (Brachydanio rerio), 5 days after fertilization. This simple sensory system consists of ten or 11 neuromasts, 15-20 sensory neurons, and about nine efferent neurons. The neuromasts are typical free neuromasts and both afferent and efferent synapses are present on hair cells within them. The sensory neurons project into a single longitudinal column of neuropil in the hindbrain. The sensory terminals appear by light microscopy to contact the dorsolateral dendrite of the ipsilateral Mauthner cell. Three types of efferent neurons are present; two types in the hindbrain and one type in the diencephalon. We provide several lines of evidence that demonstrate that these central neurons are efferent to the lateral line. We conclude from this morphology that the larval system includes all of the components of the adult system and is probably functional at this early stage. We also found that larvae have all of the efferent neurons found in adult zebrafish, while the number of neuromasts and sensory neurons will increase during subsequent development.

Brain neurons which project to the spinal cord in young larvae of the zebrafish.

A small number of brain neurons project to caudal levels of the spinal cord in the larva of the teleost Brachydanio rerio. These cells were identified in animals 6 days after fertilization by backfilling with horseradish peroxidase following transection of the cord at the level of the cloaca. In preparations with the most labeled cells a total of 30-40 were present on each side of the midline. They were located within three regions of the brainstem: the midbrain nucleus of origin of the medial longitudinal fascicle (mlf), the hindbrain reticular formation, and the hindbrain vestibular nucleus. A total of 15 classes of cells could be distinguished by soma positions, dendritic fields, and axonal pathways. For some of these classes only one or two cells were usually present on each side of the brain. Most of the labeled cells contributed axons to the mlf ipsilateral to the soma; however, the Mauthner cells and three new types of hindbrain reticulospinal reticulospinal cells have decussating axons that enter the contralateral mlf. The observed distribution of labeled reticulospinal cells is similar to that previously described for large reticular cells in adult teleosts and to the system of identified Mauthner and Müller cells in the lamprey.

Modulation of the malignant phenotype of human prostate cancer cells by N-(4-hydroxyphenyl)retinamide (4-HPR).

A long latent period of 20 to 30 years may be involved in the multistep process of carcinogenesis represented by prostatic intraepithelial neoplasia (PIN) in the prostate. It is, therefore, possible that progression to a malignant state could be blocked or reversed during this time. Retinoids not only have the ability to block steps in the process of carcinogenesis but they may also modulate or reverse some malignant characteristics of cancer cells. This study focuses on the ability of N-(4-hydroxyphenyl)-retinamide (4-HPR), a synthetic retinoid, to reverse malignant characteristics towards a normal phenotype, using the human prostate carcinoma cell line DU-145. These malignant characteristics include abnormal cell proliferation, intermediate filament expression, motility, invasion, and cell survival. Results show that 1 microM and 10 microM 4-HPR caused 31% and 96% inhibition of growth, while all-trains retinoic acid (ATRA) produced similar effects at 10 and 100 microM, making 4-HPR ten times more effective than ATRA. While DU-145 cells show strong immunostaining for vimentin, treatment with 1 microM 4-HPR for eight days caused a marked decrease in vimentin staining. This was accompanied by a change from an elongated to an epithelial cell morphology. Densitometric analysis of Western blots for vimentin showed a 53% decrease in vimentin expression in 1 microM 4-HPR treated cells. Concomitant with the decrease in vimentin expression, cell motility and invasive ability also decreased by 32% and 52%, respectively. Growth inhibition was accompanied by DNA fragmentation and apoptosis. Exposure of cells to 1 microM 4-HPR caused a marked upregulation of nuclear retinoid receptors RARalpha and a detectable expression of RARgamma. These results suggest that inhibition of growth and vimentin expression, and induction of apoptosis by 4-HPR in prostate cancer cells may occur via a receptor-mediated mechanism involving transrepression of AP-1 by retinoid receptors. We propose that vimentin may serve as a useful intermediate marker for early detection of prostate cancer in biopsy specimens and that 4-HPR may be effective in blocking several steps in prostate carcinogenesis as well as the progression of PIN to invasive carcinoma.

Hospitalization in the first year of renal replacement therapy for end-stage renal disease.

BACKGROUND: The requirement for hospitalization of patients on dialysis is likely to be a surrogate marker of age and comorbid diseases. It may also reflect the level of care delivered, and substantially increases the cost of this expensive therapy. AIM: To identify the factors most strongly associated with hospitalization. DESIGN: Prospective population study. METHODS: Data were recorded for all patients starting RRT in Scotland over one year, including the reasons for and duration of, each hospital admission during the first year of RRT. Factors most strongly associated with hospitalization were determined by Poisson regression analysis. RESULTS: Overall, 526 patients were admitted to hospital on 1668 occasions (median 3, IQR 1-4) for 13384 days (median 13, IQR 4-35). Formation of vascular access for haemodialysis (HD) was the most frequent reason for admission, followed by infections. Age, comorbidity, mode of presentation for RRT and primary renal diagnosis were all significantly associated with prolonged hospitalization. Attainment of UK Renal Association standards for urea reduction ratio and serum albumin concentration, and vascular access in the form of arterio-venous fistulae were associated with less hospitalization in patients treated with HD by 90 days. DISCUSSION: Patients in their first year of RRT have a high requirement for in-patient care, 8.6% of patient treatment days being spent in hospital. Vascular access formation, failure and complications account for a large proportion of this. Age and comorbidity prolong the time spent in hospital. As the RRT population continues to increase, with older patients and those with greater comorbidity, in-patient facilities must also expand.

Equity of renal replacement therapy utilization: a prospective population-based study.

This 1-year prospective survey assessed the incidence and characteristics of all patients starting renal replacement therapy (RRT) for end-stage renal disease in Scotland, and whether there is equity of utilization of RRT in terms of age, domicile and social circumstance. In the year studied, 104 patients per million population (533 patients) started RRT (390 per million population aged 65-75). In 23.5% the cause of ESRD could not be determined. Diabetes was the single most frequently identified cause (16%). The requirement for RRT rose with age, but over the country as a whole, patients aged over 75 years were under-represented. The majority of health boards provided RRT at a rate within 20% of the national rate. There was no difference in the median age at starting RRT between health boards. The spectrum of social deprivation of patients starting RRT was the same as that of the general population. There was no evidence that social deprivation influences acceptance on to the RRT program, although the relationship between ESRD and deprivation is complex. The utilization of RRT exceeded the minimum rate recommended by the Renal Association, although there was fluctuation between health board areas. The national requirement for resources to provide RRT is likely to rise further to care for an increasingly elderly population.

Acute renal failure requiring renal replacement therapy: incidence and outcome.

BACKGROUND: Renal replacement therapy (RRT) for acute renal failure (ARF) may be provided in many settings within the hospital. Such patients require a high level of care and often have a poor prognosis. No prospective studies have accurately defined this population, making the prediction of necessary resources and the planning of services difficult. AIM: To ascertain the incidence, causes and outcomes of acute renal failure requiring renal replacement therapy in Scotland. DESIGN: A prospective observational census of all clinical areas providing renal replacement therapy in three Scottish health boards (Grampian, Highland, Tayside). METHODS: Patients were identified by liaison with each unit providing RRT. Factors precipitating renal failure and reasons for RRT were recorded at the time of initiation. Comorbid disease burden was scored using the Charlson index. Patient status at 90 days was assessed from case-notes, contacting general practitioners where necessary. RESULTS: 375 patients per million population per year received RRT; 203 per million per year for either ARF or acute-on-chronic renal failure. 73.5% of patients receiving RRT for ARF died within 90 days, 23.5% became independent of RRT. The median duration of hospital admission was 19 days. DISCUSSION: The annual incidence of ARF requiring RRT is just over 200 per million population, almost twice that of end-stage renal disease requiring RRT. Such treatment places high demands upon health care resources.

The delivery of renal replacement therapy in Scotland--why the geographic variation?

BACKGROUND: The incidence of patients starting renal replacement therapy (RRT) for established renal failure (ERF) in Scotland has fallen from 2005 to 2011 due to a reduction in older patients starting RRT; there are significant differences between NHS Health board areas. AIM: To understand the apparent inequality in provision of RRT between NHS board areas in Scotland. DESIGN: Retrospective population analysis of Scottish renal registry (SRR) data, population statistics and quality outcomes framework summary statistics. RESULTS: The incidence of patients starting RRT for ERF in Scotland fell from 123 per million population (pmp) in 2005 to 96 pmp in 2011. The incidence of ≥75 year olds fell from 406 to 274 pmp. There are significant differences between NHS board areas when standardized for age and social deprivation. There is no relationship between the population prevalence of CKD as reported by QOF and the incidence of RRT for ERF. Those areas with high incidence rates of ≥75 year olds have higher 90-day [Spearman's rank correlation: coefficient = 0.662; P = 0.03] and 1-year [Spearman's rank correlation: coefficient = 0.776; P = 0.003] mortality rates. CONCLUSION: The significant variation in provision of RRT for ERF between Scottish NHS Board areas is not explained by age or social deprivation. There is evidence of change in practice towards RRT for patients aged ≥75 years but variation between NHS Board areas. This disparity must be further investigated to ensure equity of access to RRT for those who will benefit from it, and to non-dialytic care for those who would not.

Vascular access type and risk of mortality in a national prospective cohort of haemodialysis patients.

BACKGROUND: Central venous catheters (CVC) are a potential source of bacteraemia and have been associated with increased mortality in haemodialysis patients. We aimed to investigate the relationships between haemodialysis vascular access, taking into account changes in vascular access type during patients' lives, and cause specific mortality risk in a national cohort of dialysis patients. METHODS: Prospective cohort study including all patients receiving haemodialysis in Scotland at annual cross sectional surveys in 2009, 2010 and 2011. Data were collected through the Scottish Renal Registry and by a structured review of case records following death. Cox proportional hazards regression and multivariable logistic regression were used to model survival and risk of death from septicaemia respectively. RESULTS: Of a cohort of 2666 patients, 873 (32%) died during follow-up. After case-mix adjustment, patients using only tunnelled CVC during follow-up had a higher risk of all cause mortality across all strata of prior renal replacement therapy exposure [adjusted hazard ratio (HR): 1.83-2.08]. Case-mix adjusted risks of cardiovascular death (adjusted HR: 2.20-2.95) and infection-related death (adjusted HR: 3.10-3.63) were also higher in this group. Patients using tunnelled CVCs during follow-up and prior to death had 6.9-fold higher odds of death from septicaemia compared with those using only arteriovenous fistulae or grafts. CONCLUSION: Compared with an arteriovenous fistula or graft, sustained use of tunnelled CVCs for vascular access is associated with higher risks of all-cause, cardiovascular and infection-related mortality.

Experimental and modeling study of C5H10O2 ethyl and methyl esters.

Due to the world's over-reliance on fossil fuels there has been a developing interest in the production of renewable biofuels such as methyl and ethyl esters derived from vegetable oils and animal fats. To increase our understanding of the combustion chemistry of esters, the oxidation of methyl butanoate and ethyl propanoate, both with a molecular formula of C5H10O2, have been studied in a series of high-temperature shock tube experiments. Ignition delay times for a series of mixtures, of varying fuel/oxygen equivalence ratios (phi = 0.25-1.5), were measured behind reflected shock waves over the temperature range 1100-1670 K, and at pressures of 1.0, and 4.0 atm. It was found that ethyl propanoate was consistently faster to ignite than methyl butanoate, particularly at lower temperatures. Detailed chemical kinetic mechanisms have been assembled and used to simulate these experiments with good agreement observed. Rate of production analyses using the detailed mechanisms shows that the faster reactivity of ethyl propanoate can be explained by a six-centered unimolecular decomposition reaction with a relatively low activation energy barrier producing propanoic acid and ethylene. The elimination reaction itself is not responsible for the increased reactivity; it is the faster reactivity of the two products, propanoic acid and ethylene that leads to this behavior.

Noninvasive recording of the Mauthner neurone action potential in larval zebrafish.

We describe the identification of Mauthner (M-) cell action potentials in an intact zebrafish larva, utilizing recording electrodes located outside the fish: 1. The externally recorded spike occurs at approximately the same time, and its waveform changes with recording site in the same way, as the extracellular M-spike recorded within the central nervous system. 2. The externally recorded M-spike may be readily distinguished from other forms of neural activity. 3. The M-spike can be identified in recordings from unrestrained larvae. This finding permits the direct study of M-cell function in the freely behaving animal.

Prevalence of angiographic atherosclerotic renal artery disease and its relationship to the anatomical extent of peripheral vascular atherosclerosis.

BACKGROUND: Recognition of the possible presence of atherosclerotic renal artery disease (ARAD) is important because of its progressive nature, and because of the potential for precipitating an acute deterioration in renal function by administration of angiotensin-converting enzyme inhibitors. The aim of this study was to identify the prevalence of ARAD in patients undergoing peripheral angiography and its relationship to the extent of their peripheral vascular disease (PVD). METHODS: The reports of the 218 patients who underwent peripheral angiography to investigate PVD in one centre in a calendar year, and in whom it was possible to image the renal arteries, were analysed retrospectively. The presence of atherosclerotic disease in the renal, aortic, iliac, femoral and distal areas was recorded for each patient. RESULTS: The prevalence of ARAD was 79/218 (36.2%). The greater the number of atherosclerotic areas of the arterial tree, the higher the prevalence of ARAD. Patients with aortic disease and bilateral iliac, femoral and distal vessel disease had the highest incidence of ARAD 19/38 (50%). The incidence of ARAD in those with femoral artery atherosclerosis was significantly higher than in those without femoral artery atherosclerosis (42.1% compared with 9.7%, P=0.001 chi2). There was no significant difference in those groups with or without iliac and distal disease. None of the 11 patients with normal femoral and iliac arteries had ARAD. CONCLUSIONS: Renal artery atherosclerosis is a common occurrence in patients with PVD. If extensive PVD is recognized during aortography, a high flush should be considered to examine the renal arteries, if they are not included in the main study.

Segmental pattern of development of the hindbrain and spinal cord of the zebrafish embryo.

In the ventral hindbrain and spinal cord of zebrafish embryos, the first neurones that can be identified appear as single cells or small clusters of cells, distributed periodically at intervals equal to the length of a somite. In the hindbrain, a series of neuromeres of corresponding length is present, and the earliest neurones are located in the centres of each neuromere. Young neurones within both the hindbrain and spinal cord were identified in live embryos using Nomarski optics, and histochemically by labelling for acetylcholinesterase activity and expression of an antigen recognized by the monoclonal antibody zn-1. Among them are individually identified hindbrain reticulospinal neurones and spinal motoneurones. These observations suggest that early development in these regions of the CNS reflects a common segmental pattern. Subsequently, as more neurones differentiate, the initially similar patterning of the cells in these two regions diverges. A continuous longitudinal column of developing neurones appears in the spinal cord, whereas an alternating series of large and small clusters of neurones is present in the hindbrain.

Can we improve early mortality in patients receiving renal replacement therapy?

BACKGROUND: Approximately one in eight patients with end-stage renal disease (ESRD) die within the first three months of starting renal replacement therapy (RRT). We investigated which factors might improve this early mortality. METHODS: We performed a prospective nationwide study of all patients commencing RRT for ESRD in Scotland over one year. Patients were classified according to how they presented to start RRT, their burden of comorbid diseases, access prepared for dialysis, and duration of care by a nephrologist prior to commencing RRT. Those factors most strongly associated with death within 90 days of commencing treatment were determined by logistic regression analysis. RESULTS: Patients with an acute unexpected element to their presentation for RRT had early mortality rates between 6.0 and 8.9 times greater than those who commenced RRT electively after a period of care from a nephrologist. Patients in high and medium comorbidity risk groups had early mortality rates of 4.7 and 2.2 times greater than those in the low-risk group. Low serum albumin had a significant association with early death. Patients who progressed steadily to ESRD, who had a planned start to dialysis, and who had mature access were 3.6 times more likely to survive beyond three months than those with no access; they were, however, also younger with less comorbidity. CONCLUSIONS: The factors principally associated with early mortality are nonelective presentation for RRT, comorbid illness, and low serum albumin. Patients cared for by a nephrologist before requiring RRT who have mature access have better short-term survival than those without access. They are also younger with less comorbidity. It may be possible to improve short-term survival in this "unplanned" group if referred early to facilitate reducing cardiovascular risk factors and preparation for RRT.