Sequence variation in the capsid protein genes of human papillomavirus type 16

We have cloned and sequenced the L1 and L2 genes from human papillomavirus type 16 (HPV16) DNA-containing cervical cytology samples collected from the U.K. and Trinidad. Samples containing high copy numbers of HPV16 DNA were selected as being likely to contain fully functional virus DNA molecules in an episomal state, rather than in an integrated and possibly altered state. In comparison with the perviously published sequence of HPV16 isolated from an invasive cancer a variety of differences were detected in both L1 and L2. The pattern of changes appears to be different in samples from the two geographic regions. One of the differences (resulting in D at position 202 of the L1 protein) reported recently to be functionally important for virus particle assembly was found to occur in all the samples examined. Variations in L1 found within known immunoreactive regions or hydrophobic domains should be taken into account in design of prophylactic vaccines for HPV16 based on virus-like particles. All variations within L2 protein were found in hydrophilic domains in the carboxy-terminal half of L2. These positions were highly variable among other types of papillomavirus and are located outside the known L2 immunoreactive region. (AU)

Insulin therapy and nephropathy - abstract

There is a four-fold increase in the risk of diabetic nephropathy in patients with poor glycaemic control. Glycaemic control, soon after the diagnosis of diabetes mellitus (Type-1, IDDM), may reduce the elevated glomerular filtration rate (GFR). Recently, it has been shown that the introduction of insulin may reduce the exaggerated GFR response of newly diagnosed IDDM to infusion of amino acids to normal, and at the same time cause reversal of renal hypertrophy. Long-term studies suggest that reduction in microalbuminuria and progression to clinical nephropathy can be prevented with tight glucose control, compared to patients with lesser glucose control. In well-established nephropathy, however, there is little evidence that glycaemic control has any impact on progression to renal failure (AU)

The neurophysiological control and photosensory modulation of swimming in a cercaria, Cercaria caribbea LXXI

Scanning and transmission electron microscopy revealed that Cercaria caribbea LXXI has a small body (70 æm) with a pair of well-developed, rhabdomeric photoreceptors; and a large tail (4 mm) with six longitudinal, striated muscle bands. Muscle cells had an outer myofibrillar region adjecent to the tergument and an inner sarcoplasmic region, prolongations of which, may form sarconeural junctions. Sarcomere lengths were consistent with invertebrate fast muscle fibres. Diffuse Z-bands and the presence of numerous mitochondria are consistent with slow muscle fibres, adapted for prolonged work output. Interneuronal junctions with abundant electron-lucent and electron-dense vesicles, and neuromuscular junctions with mainly electron-lucent presynaptic vesicles were demonstrated. The neuromuscular connection between the body and tail appeared to be restricted to a single axon, implying that modulation of tail activity by the body may involve simple triggering or level setting actions, rather than complex modulation. The Falck-Hillarp Formaldehyde-Induced Fluorescence and the Gomori techniques indicated that the neuromuscular system utilizes primary catecholamines, serotonic (5-HT) and an acetylcholine-like transmitter. There were indications that 5-HT promotes activity and is held as a non-replenishable store, depleted through time within the cercarial tail. Exogenous application of a range of neuropharmacological agents, profoundly affected survivorship of the organism. Of the catecholaminergis group, propranolol (1.1-4 æM) and haloperidol (0.066-0.133 mM) most effectively reduced population half-line (T50). This was followed by the cholinergic agent, physostigmine (0.08-2.5 mM). The gamma-aminobutyric acid antagonist, picrotoxin (0.08-2.5mM), only weakly affected T50. Several drugs, especially reserpine (0.08-2.5mM), produced osmotic disturbances, leading to shortened life span. Survivorship was also dependent on pH, with a sharply defined tolerance range between pH 6 and 8. Suction electrodes recordings of electrical activity in the tail revealed spontaneous spike potentials (0.2-0.6 mV, 15-20/sec) superimposed on an underlying slow wave component. The effects of drugs on this activity were observed. 5-HT increased the amplitude and frequency of spike discharge from 0.2 mV to 0.8 mV and 15/sec to 25/sec, respectively. Physostigmine was the most effective reducer of both parameters. Indications were that cholinergic receptors in the nerve network may be excitatory nicotinic and inhibitory muscarinic. Excitatory beta adrenergic and inhibitory dopaminergic influences were observed. C. caribbea LXXI exhibited strong positive phototaxis. In response to light stimuli, the pigmented cercariae gave a maintained receptor potential with a small transient component. In the non-pigmented strain of C. caribbea LXXI, the photosensory receptor potential showed a large transient and a small maintained component. Drugs did not alter the photosensory response. A drug-induced decrease in linear translation towards a photic stimulus was therefore due to a disturbance in the neuromuscular system (AU)

Pyroglutamic aciduria of pregnancy: an index of glycine insufficiency? - abstract

We have shown that in preterm infants, and during the third trimester of pregnancy, there is a limited flow of isotopic glycine nitrogen to urea. We interpreted this as indicating that the large metabolic demand for glycine made by the rapidly growing foetus exceeded the ability of the body to produce glycine endogenously. In pyroglutamic aciduria, an inborn error of metabolism, a deficiency of glutathione synthetase results in an increased urinary excretion of pyroglutamic acid. We reasoned that limited availability of glycine, a substrate of glutathione synthetase, would restrict flow through this enzyme and might thus result in increased urinary excretion of pyroglutamic acid. We have measured the urinary excretion of pyroglutamic acid in non-pregnant and pregnant women in the 1st, 2nd and 3rd trimester. There was a relatively small dirunal variation in pyroglutamic acid/creatinine and a single urine gave a representative value. The pyroglutamic acid excretion rose progressively as pregnancy advanced and by the third trimester was over 20 times greater than in the non-pregnant women. As only a portion of the pyroglutamic acid produced is normally excreted in the urine, it is not possible to exclude the possibility that the increased excretion reflects a change in the metabolism of pyroglutamic acid unrelated to glycine metabolism. However, we have found that in four different metabolic states in which the demand for glycine is enhanced, there is an increased pyroglutamic acid excretion. These results provide support for the postulate that glycine acts as a semi-essential amino acid, and suggest that the availability of glycine may be limited as pregnancy advances (AU)

Sickle cell-Hb Lepore Boston syndrome: uncommon differential diagnosis to homozygous sickle cell disease

Two siblings assumed on the basis of clinical and hematological evidence to have homozygous sickle cell (SS) disease were found to have a mother without sickle hemoglobin. Subsequent investigation and hemoglobin structural studies indicated the diagnosis to be sickle cell-Hb Lepore Boston syndrome. This syndrome generally manifests clinically significant sickle cell disease, and this genotype should be borne in mind in apparent SS disease where a parent without sickle hemoglobin is discovered.(AU)

Nitrogen metabolism in the rat

The intermediary metabolism of 6 isotopic amino acids, 15N-aspartic acid, 15N-glutamic acid, 15N-alanine, 15N-glutamine (amide-15N), 15N-glycine and 15N-lysine (O-15N), and 15N-ammonium chloride were investigated. The aim of this study was to establish the precursor-product relationships existing between these amino acids, ammonia and urea in the amino-N pools of the major organs and tissue beds of the normal postprandial rat with the specific objective of following the movement of nitrogen to urea synthesis. It was hoped to ascertain whether glutamic acid played a central role in providing nitrogen for urea synthesis and whether there existed any relationship between 15N distribution patterns of the different isotopes and WBTP rates calculated from hepatic and renal urea-N enrichments. The method employed involved the administration of tracer quantities of the isotopes by the constant infusion technique and measuring the 15N excess of ammonia-N, glutamine amide-N, alanine-N, glutamate-N, aspartate-N and urea-N. It was found that nitrogen from 15N-alanine, 15N-aspartic acid and 15N-glutamic acid was distributed evenly in most of the amino-N pools studied. Nitrogen from the other four isotopes was distributed unevenly, preferentially to ammonia, glutamine amide and urea. 15N-glycine and 15N-lysine were only sparingly metabolised. WBPT rates obtained from urea-N enrichments were not affected by the nitrogen distribution patterns of the isotopes but by the extent to which they metabolised. WBPT rates calculated from ammonia-N enrichments were unduly affected by the extent to which each isotope contributed nitrogen to ammoniagenesis. Glutamic acid does not seem to be the precursor of both nitrogens used for urea synthesis. It supplies only one nitrogen. It is possible that urea is synthesised from an amino-N received via the glutamate to aspartate pathway and an amide-N received via the glutamine to ammonia to carbamyl phosphate pathway. Free ammonia entering the liver is first fixed as glutamine amide before being used for urea synthesis. (AU)

Renal transport and metabolism of glutamine

The experiments described in this thesis were designed to investigate the mechanism of glutamine transport and the effect of acute acidosis on glutamine transport and metabolism in the rat kidney. The experiments done with kidney slices investigated the effect of several amino acids chosen from different structural groups on uptake of glutamine. Proline (an ammino acid) and glycine (at inhibitory concentrations) were found to inhibit ammonia production and glutamine uptake significantly. When a sodium free incubation buffer was used the inhibitory effect of proline and glycine on glutamine uptake disappeared, indicating that the inhibition by proline was sodium dependent. In similar experiments with isolated kidney proximal tubules in a sodium containing buffer, proline also caused a decrease in glutamine uptake and ammonia production. Intracellular/extracellular glutamine distributions ratios in tubules however, showed a marked increase in the presence of proline. The explanation for this is not that proline shares a transport site with glutamine but rather that it is metabolised to glutamate which causes an inhibition of phosphate dependent glutaminase and hence decreased glutamine utilisation. This decreased utilisation leads to an accumulation of glutamine within the renal cell and subsequently increased intracellular/extracellular glutamine distribution ratios. The inhibitory effect of proline on glutamine uptake was much less in experiments with kidney slices from chronically acidotic rats. Isolated tubules from acutely acidotic rats showed increased intracellular/extracellular glutamine distribution ratios, increased ammonia production and glutamine uptake in tubules. When tubules from normal rats were incubated in sera from acutely acidotic rats there was a similar increase in intracellular/extracellular glutamine distribution ratios, ammonia production and glutamine uptake. The effect of the serum however, was not due to the accumulation of glutamate because glutamate levels in tubules were unchanged after incubation in sera from the acutely acidotic rats (AU)

Fasting glucose production in the smaller of twins with epinephrine-deficient hypoglycemia

The possibility that insufficient glucose production or availability of gluconeogenic substrates could account for fasting hypoglycemia was investigated in three children with epinephrine deficiency. Each had been born the smaller of discordant identical twins, and the unaffected twins served as controls. Fasting plasma glucose production was measured by constant infusion of U-[13]C-glucose under steady-state conditions and was compared with availability of potential glucose sources estimated from respiratory calorimetry and excretory nitrogen. The average rate of glucose production was 2.6 mg/kg/min in the affected twins after they became symptomatic and 2.9 mg/kg/min in the control twins after comparable fasting. Plasma alanine was lower in the affected twins during this interval (average: 0.11 mM versus 0.16 mM), but not earlier prior to decreased plasma glucose; alanine correlated with plasma glucose in a similar way in both groups (r = 0.77). Plasma urea production was 0.30 versus 0.15 mg urea N/kg/min. The calculated availability of potential gluconeogenic amino acids was 1.2 versus 0.6 mg/kg/min. Availability of glycerol, estimated from respiratory calorimetry was 0.4 mg/kg/min in both groups. In two of the twin pairs, net oxidation of carbohydrate (glycogen) was, by design, relatively small under these conditions (0.1 and 0.4 mg/kg/min in the affected and control twins, respectively). Gluconeogenesis therefore accounted for the majority of glucose production. The unaccounted remaining major gluconeogenic source is assumed to be recycled substrates from unoxidized pyruvate. Infusion of excess alanine in these two pairs increased plasma glucose and glucose production similarly in both the affected and control twins. This change was associated with an abnormally large increase in plasma alanine. In the third twin pair, net oxidation of carbohydrate was greater in the affected twin (1.8 versus 1.3 mg/kg/min) and possible glucose sources exceeded total glucose production during hypoglycemia. Earlier during fasting, net oxidation of carbohydrate in this twin was 5.8 mg/kg/min versus 3.1 mg/kg/min in the control. Plasma glucose production measured simultaneously was 4.3 versus 3.8 mg/kg/min, being less than the rate of carbohydrtae oxidation in the affected twin. It is concluded that the abnormal fasting metabolism observed in these children with decreased epinephrine was not primarily a consequence of deficient glucose production or lack of potential gluconeogenic substrates. Initial persistent oxidation of glycogen and subsequent increased utilization of protein during hypoglycemia indicate failure to conserve these limited net sources of pyruvate(AU)

In vivo studies of amino acid, water and electrolyte transport in the small intestine of normal and magnesium depleted rats

Net amino acid, water and electrolyte transport from the small intestine in vivo was studied in normal and magnesium depleted rats, by a single pass perfusion technique. The net absorption of the amino acids, alanine and lysine increased with increasing concentration of the amino acid in the perfusion solution, within the concentration range studied (10 and 50mM). In all cases alanine was absorbed faster than lysine. Dietary magnesium depletion, lasting for a period of twenty eight days did not affect the net rate of transport of these two amino acids in any of the regions of the small intestine which were studied. In general, the presence of amino acids increased the absorption of water by comparison with that from saline. However, lysine at a concentration of 50mM tended to inhibit water absorption by comparison with that observed in the presence of alanine at 50mM. Magnesium depletion did not in general affect net transport rates of water in the presence and absence of amino acids. However, when alanine at a concentration of 50mM was perfused, water absorption was inhibited in the magnesium depleted rats. The presence of amino acids did not affect the transport of sodium in any of the regions of the small intestine. However, magnesium depletion did severely inhibit sodium transport, especially in the presence of alanine at a concentration of 50mM in all three regions of the small intestine. Amino acids stimulated the absorption of chloride by all segments of the small intestine by comparison with that from saline alone. Magnesium depletion however, significantly reduced chloride ion absorption by all three segments. Good correlations were found between the transport rates of sodium and chloride, sodium and water, and total solute and water in all three regions of the small intestine (AU)

The in vivo measurement of protein synthesis

Methods of measuring in vivo protein synthesis are briefly reviewed. Methods involving incorporation of label into protein are more appropriate for mixed proteins. The major difficulty is the definition of the precursor for protein synthesis. The only data available on the effect of infection on protein synthesis are open to criticism on the grounds that the precursor pool was not sampled. (AU)

The measurement of rates of protein turnover, synthesis, and breakdown in man and the effect of nutritional status and surgical injury

Current concepts of protein turnover, synthesis and breakdown in man are reviewed. Emphasis is placed on the distinction between protein breakdown, by which constituent amino acids are released, and amino acid catabolism, which leads to the formation of excretory products, mainly urea and carbon dioxide. Methods of measuring overall rates of turnover, synthesis, and breakdown of body protein in man by the continuous or intermittent administration of radioactive or stable isotopes of amino acids are reviewed. Data from the published literature and from ongoing studies reveal that the overall rate of protein synthesis declines with age. There is close agreement between results obtained with different isotopes in normal adults. Malnourished infants have significantly lower rate of protein turnover, synthesis and breakdown before than after recovery. These finding may represent another example of metabolic adaptation in the malnourished child. After surgical trauma in adults the rate of protein synthesis is significantly reduced and neither protein turnover nor breakdown shows a significant change from preoperative values. It is concluded from these studies that the negative nitrogen balance found after injury is accounted for by reduction in protein synthesis and that protein breakdown remains unchanged after orthopedic operations. The relevance of these findings to the changes in nitrogen metabolism seen in infection is discussed (AU)

Haemoglobin Spanish Town alpha27 Glu replaced by Val (B8)

A new alpha chain variant Hb Spanish Town, a27 Glutamic acid-Valine, awas detected in the cord blood of a Jamaica Negro infant. In the mother the adult component (a2 Spanish TownB2) has an electrophoretic mobility between haemoglobins S and F at alkaline pH and measures 11.0-12.0 percent of the total haemoglobin. (Summary)