Maternal hemodynamics, fetal biometry and Doppler indices in pregnancies followed up for suspected fetal growth restriction.

OBJECTIVES: To assess whether, in a cohort of patients with a small-for-gestational-age (SGA) fetus with estimated fetal weight ≤ 10th percentile, maternal hemodynamics, fetal biometry and Doppler indices at presentation can predict the subsequent development of an abnormal fetal Doppler index or delivery of a baby with birth weight < 3rd percentile. METHODS: This was a prospective observational cohort study conducted at a specialist clinic for the management of pregnancies with a SGA fetus at King's College Hospital, London, UK. The study population comprised 86 singleton pregnancies with a SGA fetus, presenting at a median gestational age of 32 (range, 26-35) weeks. We measured maternal cardiac function using a non-invasive transthoracic bioreactance monitor, as well as mean arterial pressure, fetal biometry, and umbilical artery (UA), fetal middle cerebral artery (MCA) and uterine artery (UtA) pulsatility indices (PI), and the deepest vertical pool of amniotic fluid. Z-scores of these variables were calculated based on reported reference ranges and the values were compared between pregnancies with evidence of an abnormal fetal Doppler index at presentation (Group 1), those that had developed an abnormal Doppler index at a subsequent visit (Group 2) and those that did not develop an abnormal Doppler index throughout pregnancy (Group 3). Abnormal fetal Doppler was defined as UA-PI > 95th percentile and/or MCA-PI < 5th percentile. Differences in measured variables at presentation were also compared between pregnancies delivering a baby with birth weight < 3rd percentile and those delivering a baby with birth weight ≥ 3rd percentile. Multivariate logistic regression analysis was used to determine significant predictors of birth weight < 3rd percentile and evolution from normal to abnormal fetal Doppler. RESULTS: In the study population, 14 (16%) cases were in Group 1, 19 (22%) in Group 2 and 53 (62%) in Group 3. Birth weight was < 3rd percentile in 39 (45%) cases and ≥ 3rd percentile in 47 (55%). There was decreased cardiac output and stroke volume and increased peripheral vascular resistance compared with a normal population, and the deviations from normal were most marked in Group 1. Pregnancies with birth weight < 3rd percentile, compared with those with birth weight ≥ 3rd percentile, had greater deviations from normal in fetal biometry, maternal cardiac output, stroke volume, heart rate, peripheral vascular resistance and UtA-PI. Multivariate logistic regression analysis demonstrated that, in the prediction of birth weight < 3rd percentile, maternal hemodynamic profile provided significant improvement to the prediction provided by maternal demographics, fetal biometry, UtA-PI, UA-PI and MCA-PI (difference between areas under receiver-operating characteristics curves, 0.18 (95% CI, 0.06-0.29); P = 0.002). In contrast, there was no significant independent contribution from maternal hemodynamics in the prediction of the subsequent development of abnormal fetal Doppler. CONCLUSIONS: In pregnancies with a SGA fetus, there is decreased maternal cardiac output and stroke volume and increased peripheral vascular resistance, and the deviations from normal are most marked in cases of redistribution in the fetal circulation and reduced amniotic fluid volume. Copyright © 2018 ISUOG. Published by John Wiley & Sons Ltd.

Molecular tagging of a senescence gene by introgression mapping of a stay-green mutation from Festuca pratensis.

* Intergeneric hybrids between Lolium multiflorum and Festuca pratensis (Lm/Fp) and their derivatives exhibit a unique combination of genetic and cytogenetic characteristics: chromosomes undergo a high frequency of homoeologous recombination at meiosis; the chromosomes of the two species can easily be discriminated by genomic in situ hybridization (GISH); recombination occurs along the entire length of homoeologous bivalents; a high frequency of marker polymorphism is observed between the two species. * This combination of characters has been used to transfer and isolate a F. pratensis chromosome segment carrying a mutant 'stay-green' gene conferring a disrupted leaf senescence phenotype into L. multiflorum. * The genetic location within the introgressed F. pratensis segment of the senescence gene has been mapped using amplified fragment length polymorphisms (AFLPs), and F. pratensis-specific AFLP markers closely flanking the green gene have been cloned. * The use of these cloned sequences as markers for the stay-green locus in marker-assisted selection programmes has been tested. The potential application of Lm/Fp introgressions as a tool for the map-based cloning of introgressed Fp genes is discussed.

Systemic lupus erythematosus in Trinidadian children.

Thirty-three children with a diagnosis of systemic lupus erythematosus (SLE) were studied. At diagnosis, 29 of them (88%) were aged between 10 and 17 years and the other four (12%) between 5 and 9 years. The majority were girls (28, 82%) and the male:female ratio was 1:6.6. Children of East Indian and mixed racial origin formed the largest groups (37 and 39%, respectively) and mortality was higher in these two groups. The most common symptoms at diagnosis were: fever for > 1 week (75.8%), musculoskeletal symptoms (arthralgia, arthritis and myalgia (69.7%) and renal involvement (63.6%). Malar and discoid rashes were common, 39 and 37%, respectively. Central nervous system involvement at presentation was a rare but important cause of mortality. The mortality rate during follow-up was high at 39.3% and the commonest cause of death was renal failure. Childhood SLE is uncommon in Trinidad and Tobago. Diagnosis is often delayed because of the protean and non-specific manifestations. This study reports a higher prevalence, a more severe course and greater mortality in children of East Indian and mixed descent than in children of African origin. It also shows that the symptomatology at first presentation is consistent with other studies and should be recognised early. Early diagnosis and prompt and appropriate management are essential in order to reduce the high mortality still associated with SLE.

Systemic lupus erythematosus in Trinidadian children

Thirty-three children with a diagnosis of systemic lupus erythematosus (SLE) were studied. At diagnosis, 29 of them (88%) were aged between 10 and 17 years and the other four (12%) between 5 and 9 years. The majority were girls (28, 82%) and the male:female ratio was 1:6.6. Children of East Indian and mixed racial origin formed the largest groups (37 and 39%, respectively) and mortality was higher in these two groups. The most common symptoms at diagnosis were: fever for > 1 week (75.8%), musculoskeletal symptoms (arthralgia, arthritis and myalgia (69.7%) and renal involvement (63.6%). Malar and discoid rashes were common, 39 and 37%, respectively. Central nervous system involvement at presentation was a rare but important cause of mortality. The mortality rate during follow-up was high at 39.3% and the commonest cause of death was renal failure. Childhood SLE is uncommon in Trinidad and Tobago. Diagnosis is often delayed because of the protean and non-specific manifestations. This study reports a higher prevalence, a more severe course and greater mortality in children of East Indian and mixed descent than in children of African origin. It also shows that the symptomatology at first presentation is consistent with other studies and should be recognised early. Early diagnosis and prompt and appropriate management are essential in order to reduce the high mortality still associated with SLE.

The actions of anandamide on rat superficial medullary dorsal horn neurons in vitro.

Whole-cell patch-clamp recordings were made from neurons in the trigeminal nucleus caudalis and trigeminal ganglion, in vitro, to investigate the cellular actions of the endogenous cannabinoid, anandamide. Anandamide has been shown to act through both the cannabinoid receptor 1 (CB1) and the vanilloid receptor 1 (VR1). Anandamide (30 microM) caused a 54 % increase in the rate of miniature excitatory post-synaptic currents (mEPSCs), without affecting their amplitude. The effect of anandamide was blocked by the VR1 antagonist capsazepine (20 microM), but not by the CB1-specific antagonist AM251 (3 microM). Application of the VR1 receptor agonist capsaicin (300 nM) caused a 4200 % increase in the mEPSC rate. In dissociated trigeminal ganglion neurons, both anandamide and capsaicin caused an outward current in neurons that were voltage clamped at +40 mV. The maximal outward current produced by anandamide (EC50, 10 microM) was 45 % of that produced by capsaicin (10 microM). Co-application of the VR1 antagonist capsazepine (30 microM) completely reversed the effects of both capsaicin and anandamide. The anandamide transport inhibitor, AM404 (30 microM) caused a 40 % increase in mEPSC rate in the slice preparation and an outward current in dissociated neurons. The latter current was reversed by the VR1 antagonist iodoresiniferatoxin (1 microM). The fatty acid amide hydrolase (FAAH) inhibitors phenylmethylsulfonyl fluoride (PMSF) (20 microM) and OL53 (1 microM) did not enhance the effect of anandamide in either the slice or dissociated neuron preparations. These results suggest that within the superficial medullary dorsal horn, anandamide (30 microM) acts presynaptically to enhance the release of glutamate via activation of the VR1 receptor.

A demonstration of a 1:1 correspondence between chiasma frequency and recombination using a Lolium perenne/Festuca pratensis substitution.

A single chromosome of the grass species Festuca pratensis has been introgressed into Lolium perenne to produce a diploid monosomic substitution line 2n = 2x = 14. The chromatin of F. pratensis and L. perenne can be distinguished by genomic in situ hybridization (GISH), and it is therefore possible to visualize the substituted F. pratensis chromosome in the L. perenne background and to study chiasma formation in a single marked bivalent. Recombination occurs freely in the F. pratensis/L. perenne bivalent, and chiasma frequency counts give a predicted map length for this bivalent of 76 cM. The substituted F. pratensis chromosome was also mapped with 104 EcoRI/Tru91 and HindIII/Tru91 amplified fragment length polymorphisms (AFLPs), generating a marker map of 81 cM. This map length is almost identical to the map length of 76 cM predicted from the chiasma frequency data. The work demonstrates a 1:1 correspondence between chiasma frequency and recombination and, in addition, the absence of chromatid interference across the Festuca and Lolium centromeres.

Physical and genetic mapping in the grasses Lolium perenne and Festuca pratensis.

A single chromosome of the grass species Festuca pratensis has been introgressed into Lolium perenne to produce a diploid monosomic substitution line 2n = 2x = 14. In this line recombination occurs throughout the length of the F. pratensis/L. perenne bivalent. The F. pratensis chromosome and recombinants between it and its L. perenne homeologue can be visualized using genomic in situ hybridization (GISH). GISH junctions represent the physical locations of sites of recombination, enabling a range of recombinant chromosomes to be used for physical mapping of the introgressed F. pratensis chromosome. The physical map, in conjunction with a genetic map composed of 104 F. pratensis-specific amplified fragment length polymorphisms (AFLPs), demonstrated: (1) the first large-scale analysis of the physical distribution of AFLPs; (2) variation in the relationship between genetic and physical distance from one part of the F. pratensis chromosome to another (e.g., variation was observed between and within chromosome arms); (3) that nucleolar organizer regions (NORs) and centromeres greatly reduce recombination; (4) that coding sequences are present close to the centromere and NORs in areas of low recombination in plant species with large genomes; and (5) apparent complete synteny between the F. pratensis chromosome and rice chromosome 1.

An outbreak of measles in a rural Queensland town in 1997; an opportunity to assess vaccine effectiveness.

This report describes a measles outbreak in a rural town in south-east Queensland and presents the results of a vaccine effectiveness (VE) study performed during this outbreak. It is important to assess the effectiveness of a vaccine in an outbreak to determine if the outbreak is due to failure of the vaccine or failure to vaccinate. There were 44 cases of measles amongst local residents, which represents a notification rate of 396.7 per 100,000 population. Case investigations identified a group of people who had been exposed to measles at a seminar. The attack rate for the seminar cohort was 18% (11/61). This presented an opportunity to conduct a VE study using data about children aged less than 16 years who attended the seminar. In this cohort of 23 attendees, all 7 children who had not received any measles vaccinations became cases whilst the 6 who were fully vaccinated for their age according to NHMRC guidelines were protected from measles illness. Although there were insufficient fully vaccinated cohort members to reliably estimate VE for this group, the vaccine was 84.6% (95% CI: 15.0-99.7%) effective for those who had received at least one validated dose of vaccine. Despite the sample size limitations, the results support the view that failure to vaccinate rather than vaccine failure contributed to the high infection rate in the seminar cohort.

Middle ear effusion: rate and risk factors in Australian children attending day care.

There have been no previous longitudinal studies of otitis media conducted in non-Aboriginal Australian children. This paper describes the rate and risk factors for middle ear effusion (MEE) in children attending day care in Darwin, Australia. A prospective cohort study of 252 children under 4 years was conducted in 9 day care centres over 12 fortnights between 24 March and 15 September 1997. Tympanometry was conducted fortnightly and multivariate analysis used to determine risk factors predicting MEE. The outcome of interest was the rate of type B tympanograms per child detected in either ear at fortnightly examinations. After adjusting for clustering by child, MEE was detected on average 4.4 times in 12 fortnights (37% of all examinations conducted). Risk factors associated with presence of effusion were younger age, a family history of ear infection, previous grommets (tympanostomy tubes), ethnicity and the day care centre attended. A history of wheeze appeared protective. These effects were modest (RR 0.57-1.70). Middle ear effusion is very common in children attending day care in Darwin. This has clinical importance, since MEE during early childhood may affect optimal hearing, learning and speech development. There is little scope for modification for many of the risk factors for MEE predicted by this model. Further study of the day care environment is warranted.

A retrospective search for lyssavirus in humans in the Northern Territory.

BACKGROUND: Following the 1996 discovery of a rabies-like lyssavirus in Australian flying foxes, it was unclear whether this was a new epizootic or an unrecognised, previously existing disease. OBJECTIVE: To review cases of unexplained encephalitis in the Northern Territory (NT) to test available clinical specimens for lyssavirus and survey the use of diagnostic tests by clinicians. METHODS: The NT hospital morbidity database was searched from January 1992 to September 1996 for all Royal Darwin Hospital (RDH) cases with an ICD-9 code encompassing encephalitis or viral meningitis. Final diagnoses were determined by hospital record review. For cases of unexplained encephalitis, we assessed the use of diagnostic tests and located clinical specimens for testing for lyssavirus-specific inclusion bodies via immunohistochemistry, immunofluorescence and reverse-transcriptase polymerase chain reaction (RT-PCR). RESULTS: Encephalitis occurred in 34/154 (22%) cases located by the search; 53% (18/34) of encephalitis cases were unexplained. Of these, 24% had no serology performed and 47% had no blood cultures taken. Four (22%) died and two had autopsies. These were the only two cases with clinical specimens available for testing. They were negative for lyssavirus. None of the 71 cases coded as viral meningitis had unexplained encephalitis. CONCLUSION: There was a considerable proportion of unexplained illness among NT cases of encephalitis. IMPLICATIONS: Clinicians should test for lyssavirus in patients with encephalitic symptoms and a postmortem should be sought where death is unexplained. Specimens should be stored to enable testing for emerging infectious diseases.

Investigation of a cluster of Staphylococcus aureus invasive infection in the top end of the Northern Territory.

INTRODUCTION: Staphylococcus aureus invasive infection remains a serious condition associated with considerable morbidity and mortality. Following notification of five cases at Royal Darwin Hospital (RDH), we searched for related cases, determined their epidemiological characteristics and attempted to identify the source of this apparent cluster. METHODS: We reviewed RDH microbiology records between June 1996 and April 1997 for S. aureus isolates with similar antibiograms to notified cases. We used antibiotic resistance patterns, bacteriophage typing and two molecular typing techniques to subtype implicated isolates. Hospital records were reviewed for admission details and associated costs were estimated. RESULTS: Fifty-four cluster-related isolates occurred in 47 separate presentations. The peak incidence was in the wet season. The most important risk factor for staphylococcal invasive infection was the presence of skin sores/scabies in 17/54 cases (31%), followed by intravascular line use in 14/54 (26%), open trauma in 11/54 (20%), underlying end stage renal failure and alcoholism each in ten of 54 (18%). The mean admission length was 30 days and antibiotics were given for an average of 23 days. Death due to S. aureus infection occurred in eight of 47 (17%) presentations. S. aureus pneumonia was community acquired in 12/13 patients (92%) and six of 13 (46%) died. Ten of 13 (80%) pneumonia patients had at least one other focus of S. aureus infection. The cost of antibiotics and hospital bed per presentation was approximately $16,000. Presentations with skin sores/scabies cost considerably more ($31,000). No common epidemiologic features were found for community or hospital acquired cases. CONCLUSION: Considerable mortality and cost was attributable to cases of S. aureus invasive infection during this cluster; particularly those with community acquired pneumonia or skin sores/scabies. Staphylococcal antibiotic cover should be considered early for unwell patients presenting to hospital with pneumonia and other signs of potential S. aureus infection. It is appropriate to target public health efforts to prevent skin sores and to provide adequate treatment when they occur.

Blood pressure and calcium intake are related to bone density in adult males.

Based on the premise that elevated blood pressure and low bone mass have both been associated with poor Ca nutriture and disturbances in Ca metabolism, a cross-sectional study was employed to determine if blood pressure and dietary Ca intake were significantly related to bone mass. Forty-seven men between 24-77 years of age with blood pressure values ranging from normal to mildly elevated comprised the study group. Blood pressure was measured with a random-zero sphygmomanometer. Bone mineral content (BMC) and density (BMD) of the hip, spine and total body were measured with dual-photon absorptiometry. Dietary intake and physical activity were also assessed. Multiple linear regression analysis was used for statistical analysis. After adjusting for known confounding variables (age, BMI, Ca intake, and others) diastolic blood pressure was negatively related to BMC (P < or = 0.05) and BMD (P < or = 0.01) of the total body, trochanteric region (P < 0.01) and Ward's triangle (P < 0.05), and to BMC of the femoral neck (P < 0.05) and lumbar spine, although the latter was just shy of statistical significance (P = 0.058). Systolic blood pressure was negatively related to trochanteric BMD (P = 0.04) and BMC (P = 0.06). Ca intake was positively related to total body BMD (P = 0.005), and BMC of the lumbar spine (P = 0.05). In this population of men, Ca intake was a positive predictor, and blood pressure was a negative predictor of regional measures of bone mass. These findings support the concept that independent of age, BMI and Ca intake, elevated blood pressure varies indirectly with bone mass and density, known predictors of osteoporotic fractures. Future studies are needed to determine whether elevated blood pressure is causally related to the development of low bone mass, and what role dietary Ca plays in that pathway.

Involvement of two isoforms of SNAP-25 in the expression of long-term potentiation in the rat hippocampus.

Induction of long-term potentiation (LTP) in the hippocampus is associated with changes in expression of a variety of different proteins and is thought to be the mechanism which underlies synaptic plasticity. The 25 kDa synaptosomal-associated protein (SNAP-25) is a presynaptic protein which is involved in neurotransmitter exocytosis at the nerve terminal. Two isoforms of SNAP-25 have so far been identified (a and b) which differ in their distribution and developmental regulation. Using in situ hybridization, we demonstrated that the mRNA levels of the two isoforms of this protein are increased 2 h after the induction of LTP in granule cells of the dentate gyrus following high frequency stimulation of the perforant path in vivo. These observations further demonstrate the involvement of both isoforms of SNAP-25 in functional synaptic plasticity, although their exact roles have yet to be fully determined.

Increased expression of dendritic mRNA following the induction of long-term potentiation.

A small number of mRNAs, including Ca2+/calmodulin-dependent protein kinase II alpha-subunit (CamKIIalpha) mRNA and microtubule-associated protein 2 (MAP2) mRNA, are present in the dendrites of neurones as well as in the cell bodies. We show here that the induction of long-term potentiation (LTP) in the hippocampal perforant path/granule cell synapses in anaesthetised rats is associated with increased levels of CamKIIalpha mRNA and MAP2 mRNA in the granule cell dendrites after 2 h. Similarly, induction of LTP in the Schaffer collateral/CA1 pyramidal cell synapses in hippocampal slices maintained in vitro also results in elevated dendritic levels of CamKIIalpha mRNA and MAP2 mRNA 2 h later. In both models, the levels of various other mRNA species restricted to the cell body region were unaffected by the induction of LTP. Increased expression of dendritic CamKIIalpha mRNA and MAP2 mRNA appears to be a general feature of hippocampal plasticity, since it occurs following LTP induction in both the dentate gyrus and the CA1 region. The elevation of mRNA levels in a restricted region close to the afferent synapses would allow a highly-localised enhancement of the synthesis of the corresponding proteins, providing an elegant mechanism for protein-synthesis-dependent synaptic plasticity to maintain a high degree of anatomical specificity.

Two Afro-Trinidadian siblings with end-stage renal disease.

The case histories of two Afro-Trinidadian brothers aged 8 and 11 years who developed end-stage renal disease (ESRD) are presented. Neither had had cause in the past to seek medical attention for any renal-related illness. At presentation both had anaemia, growth failure and other clinical and laboratory evidence of ESRD. Kidney histology in one child was consistent with familial juvenile nephronophthisis (NPH). This is a common cause of ESRD in children in other countries but it has not been recognized previously in Trinidadian and other West Indian children, and should be considered as a possible aetiology in West Indian children presenting with renal failure.

Evidence for the presence of smooth muscle alpha-actin within pericytes of the renal medulla.

This study was designed to determine whether smooth muscle alpha-actin mRNA and smooth muscle alpha-actin contractile protein elements were present within the renal medullary pericytes. Extraction of total RNA from microdissected outer medullary descending vasa recta allowed for the detection of smooth muscle alpha-actin mRNA expression using reverse transcription-polymerase chain reaction (RT-PCR). Expression of smooth muscle alpha-actin was specific to the descending vasa recta and not a result of tubular contamination because RT-PCR amplification of the vasopressin V2 receptor, which is a specific tubular marker, did not occur. To determine the exact cell type(s) that translate the mRNA into protein, we performed immunohistochemistry on the renal outer and inner medulla using a monoclonal smooth muscle alpha-actin antibody, whose specificity was determined by immunoblot analysis. Smooth muscle alpha-actin protein was found selectively within the pericytes surrounding the descending vasa recta from the outer and inner medullary tissue sections. This study demonstrates that the pericytes alone that surround the descending vasa recta within the outer and inner medulla contain smooth muscle alpha-actin mRNA and protein and are therefore the site of the contractile elements that could play a vasomodulatory role in the control of renal medullary blood flow and its distribution within the renal medulla.

Long-term potentiation in perforant path/granule cell synapses is associated with a post-synaptic induction of proenkephalin gene expression.

Enkephalin peptides released from hippocampal mossy fibres lower the threshold for the generation of long-term potentiation (LTP) at the mossy fibre synapses. High frequency stimulation of the hippocampal dentate gyrus, sufficient to induce mossy fibre LTP, is associated with increased expression of the proenkephalin gene in the granule cells. We show here that a similar elevation in proenkephalin mRNA levels is observed, in anaesthetised rats, following stimulation of the perforant path sufficient to induce LTP in the perforant path/granule cell synapses. This strengthens the evidence implicating granule cell enkephalins as mediators of functional plasticity in the hippocampus. Furthermore. the results hint at a form of 'domino plasticity', where potentiation of transmission at the perforant path/granule cell synapses is subsequently followed by an enkephalin-mediated potentiation of transmission at the mossy fibre synapses.

Changes in hippocampal gene expression associated with the induction of long-term potentiation.

The expression of four genes: zif/268, c-fos, tubulin and alpha Ca2+/calmodulin-dependent protein kinase II (alpha CAMKII) was studied following the induction of LTP in Schaffer collateral CA1 neurone synapses in rat hippocampal slices maintained in vitro. Levels of c-fos mRNA and tubulin (T26) mRNA in area CA1 were unchanged after induction of LTP, however, zif/268 and alpha CAMKII mRNA levels showed a significant increase compared to non-potentiated controls. It is possible, therefore, to measure changes in gene expression using in situ hybridisation following induction of LTP in vitro and these results strengthen the theory that zif/268 and alpha CAMKII are involved in some aspect of the induction or maintenance of hippocampal LTP.