Genetic analysis of glutamate receptors in Drosophila reveals a retrograde signal regulating presynaptic transmitter release.

Postsynaptic sensitivity to glutamate was genetically manipulated at the Drosophila neuromuscular junction (NMJ) to test whether postsynaptic activity can regulate presynaptic function during development. We cloned the gene encoding a second muscle-specific glutamate receptor, DGluRIIB, which is closely related to the previously identified DGluRIIA and located adjacent to it in the genome. Mutations that eliminate DGluRIIA (but not DGluRIIB) or transgenic constructs that increase DGluRIIA expression were generated. When DGluRIIA is missing, the response of the muscle to a single vesicle of transmitter is substantially decreased. However, the response of the muscle to nerve stimulation is normal because quantal content is significantly increased. Thus, a decrease in postsynaptic receptors leads to an increase in presynaptic transmitter release, indicating that postsynaptic activity controls a retrograde signal that regulates presynaptic function.

Postsynaptic PKA controls quantal size and reveals a retrograde signal that regulates presynaptic transmitter release in Drosophila.

Two distinct mechanisms regulate synaptic efficacy at the Drosophila neuromuscular junction (NMJ): a PKA-dependent modulation of quantal size and a retrograde regulation of presynaptic release. Postsynaptic expression of a constitutively active PKA catalytic subunit decreases quantal size, whereas overexpression of a mutant PKA regulatory subunit (inhibiting PKA activity) increases quantal size. Increased PKA activity also decreases the response to direct iontophoresis of glutamate onto postsynaptic receptors. The PKA-dependent modulation of quantal size requires the presence of the muscle-specific glutamate receptor DGluRIIA, since PKA-dependent modulation of quantal size is lost in homozygous viable DGluRIIA- mutants. Furthermore, elevated postsynaptic PKA reduces the quantal amplitude and the time constant of miniature excitatory junctional potential (mEJP) decay to values that are nearly identical to those observed in DGluRIIA mutants. The PKA-dependent reduction in quantal size is accompanied developmentally by an increase in presynaptic quantal content, indicating the presence of a retrograde signal that regulates presynaptic release.

Canine intraspinal meningiomas: imaging features, histopathologic classification, and long-term outcome in 34 dogs.

BACKGROUND: Meningioma is the most common primary intraspinal nervous system tumor in dogs. Clinical findings, clinicopathologic data, and treatment of these tumors have been reported sporadically, but little information is available regarding cerebrospinal fluid (CSF) analysis, histologic tumor grade, or efficacy of radiation therapy as an adjunct to cytoreductive surgery. ANIMALS: Dogs with histologically confirmed intraspinal meningiomas (n = 34). METHODS: A retrospective study of dogs with intraspinal meningiomas between 1984 and 2006 was carried out. Signalment, historical information, physical examination, clinicopathologic data, radiation therapy protocols, surgery reports, and all available images were reviewed. All tumors were histologically classified and graded as defined by the international World Health Organization classification scheme for central nervous system tumors. RESULTS: Intraspinal mengiomas in dogs are most common in the cervical spinal cord but can be found throughout the neuraxis. Location is correlated with histologic grade, with grade I tumors more likely to be in the cervical region than grade II tumors. Myelography generally shows an intradural extramedullary compressive lesion. On magnetic resonance imaging, the masses are strongly and uniformly contrast enhancing and a dural tail often is present. CSF analysis usually shows increased protein concentration with mild to moderate mixed pleocytosis. Surgical resection is an effective means of improving neurologic status, and adjunctive radiation therapy may lead to an improved outcome. CONCLUSIONS AND CLINICAL IMPORTANCE: Biopsy is necessary for definitive diagnosis, but imaging and CSF analysis can suggest a diagnosis of meningioma. Treatment of meningiomas with surgery and radiation therapy can result in a fair to excellent prognosis.

Distinguishing between natural and aquaculture-derived sediment concentrations of heavy metals in the Broughton Archipelago, British Columbia.

Marine sediment samples were collected in the Broughton Archipelago, British Columbia, to assess the use of a geochemical normalization technique in the identification of a chemical tracer of aquaculture waste material. Zinc and copper were suggested as tracers of feed pellets, while copper was considered an indicator of anti-foulant agents used on netpen systems. The sediment samples were analyzed for carbon, nitrogen, organic matter, water, trace-element, and free sulfide concentrations, and sediment grain-size distribution. Sediment texture analysis revealed a wide range of substrate types from sand to silty loam categories. Strong relationships between sediment texture, sediment porosity, and organic content were observed across both near-field and far-field stations. Excess zinc and copper sediment concentrations, identified using a lithium-normalization technique, were restricted to near-field sampling stations (0 and 30m from netpen systems). The relationships between these metal tracers and organic content and sulfur concentrations were explored to account for variations in sediment concentrations of zinc and copper.

The use of meiofauna as an indicator of benthic organic enrichment associated with salmonid aquaculture.

Sediment chemistry and meiofaunal samples were collected in the Broughton Archipelago, British Columbia, to examine the relationship between sediment variables and meiofauna diversity across near- and far-field gradients surrounding several finfish aquaculture operations. The sediment variables examined consisted of free sulfide concentrations, redox potential (E(NHE)), organic content, and sediment grain size. A strong trend between sediment texture and organic content was observed across all sampling locations, which reflected a wide range of sediment types (sand-silt) that exist within the Broughton Archipelago. The abundance of certain meiofaunal groups (kinorhynchs, crustaceans, polychaetes) declined in an asymptotic fashion with increasing free sulfide concentrations, an indicator of benthic organic enrichment. Within these relationships, low meiofaunal abundances occurred in fine sediments associated with higher organic contents. Although other groups (nematodes, foraminifera) showed a slight decline in abundance with increasing organic enrichment, a high amount of variability in abundance was observed at high free sulfide concentrations, rendering these groups not suitable as indicators of organic enrichment. This study reports both horizontal and vertical trends in meiofaunal distributions relative to free sulfide concentrations. The abundance of nematode and crustacean taxa decreased with sediment depth as well as increasing sulfide concentration, while polycheate abundance increased with increasing free sulfide concentration. The ratio of nematodes to copepods was also shown to represent the degree of organic loading associated with aquaculture operations and its application as an indicator of benthic impact is discussed.

ATP overflow from the mouse isolated vas deferens.

1. A modified form of the highly specific luciferin-luciferase assay was used to measure the overflow of adenosine 5'-triphosphate (ATP) from the field-stimulated mouse vas deferens in vitro. Precise timing of the stimulation, superfusate collection and assay minimized ATP degradation before assay, offering the opportunity for quantitative studies. 2. Stimulation with between 50 and 200 supramaximal pulses at 5 Hz increased ATP overflow by between 7 and 28 times over basal. 3. ATP overflow increased steadily with increasing numbers of stimuli from 50 to 200 pulses. Increasing the frequency of stimulation initially had no effect, but above 10 Hz there was increased overflow, suggesting an interaction between facilitated release and the rate of degradation by extracellular ATPases. 4. Contractions of the vas induced by exogenous (-)-phenylephrine produced only a small increase in ATP overflow, suggesting that the stimulation induced increase in ATP overflow is mostly pre-junctional in origin, though tetrodotoxin eliminated stimulation-induced overflow only in a proportion of preparations.

Interactions between the effects of yohimbine, clonidine and [Ca]o on the electrical response of the mouse vas deferens.

Excitatory junction potentials (e.j.ps) were recorded from mouse vas deferens and resolved into families of 'discrete events' (d.es) reflecting intermittent release of packets of transmitter from one or a few sites. Within families d.es vary in amplitude between a few preferred values unaffected by any treatments used in these experiments. As [Ca]o is raised from 1.1 to 4.0 mM there is a rise in d.e. amplitude due to an increase in the frequency of large events and a decrease in that of small. At all [Ca]o clonidine reduces d.e. amplitude by increasing failures and small events and decreasing large events. Yohimbine has opposite effects. Both drug effects are concentration-dependent in the range 5 X 10(-9) - 10(-6)M. As [Ca]o is raised from 1.1 to 4.0 mM, and therefore more natural agonist is released, clonidine becomes more effective at altering d.e. amplitude whereas yohimbine becomes less so. With very low frequency stimulation yohimbine elevates e.j.p. amplitude only if [Ca]o is below 1.6 mM. These results are not easily compatible with the notion that yohimbine breaks a 'negative feedback' control of transmitter release.

A study of the actions of P1-purinoceptor agonists and antagonists in the mouse vas deferens in vitro.

1. We have examined the effects of purinoceptor agonists and antagonists on the mechanical 'twitch' response, excitatory junction potential (e.j.p.) amplitude and [3H]-noradrenaline overflow in the mouse vas deferens. 2. The agonist profile for inhibition of the mechanical response was N6-([R]-2-phenylisopropyl)adenosine (L-PIA) congruent to N6-cyclohexyladenosine (CHA) greater than 5' N-ethylcarboxamido-adenosine (NECA) greater than 2-chloroadenosine (2ClA) congruent N6-([S]-2-phenylisopropyl)adenosine (D-PIA). 3. The P1-purinoceptor agonists inhibited the e.j.p. with an agonist profile of CHA greater than L-PIA congruent to NECA greater than 2ClA. 4. 2ClA inhibited [3H]-noradrenaline overflow with an EC50 of 1.2 microM which was not significantly different from the values for inhibition of the e.j.p. and the mechanical response. 5. The inhibitory action of 2ClA on the mechanical response was antagonized by 5 microM 8-phenyltheophylline (8-PT). However, neither blockade of P1-purinoceptors by 8-PT nor increasing the rate of degradation of endogenous adenosine by addition of adenosine deaminase had any effect on the mechanical response per se. 8-PT (5 microM) also failed to alter the e.j.p. amplitude or [3H]-noradrenaline overflow. 6. These results indicate that there are P1-purinoceptors present on sympathetic nerve terminals of the mouse vas deferens which are more like A1- than A2-receptors, but may be better classified as being of the A3-subtype (Ribeiro & Sebastiao, 1986). These receptors are not normally involved in the feedback regulation of transmitter release in this tissue.

Relations between the development of patterns of sleeping heart rate and body temperature in infants.

Overnight patterns of rectal temperature and heart rate were recorded from 119 normal infants at weekly intervals from 7 to about 16 weeks of age. All data were collected in the infants' own homes. As previously reported, different infants developed an adult-like night time rectal temperature pattern abruptly at different ages. When heart rate data were collated by age, there was an apparently gradual fall in sleeping heart rate from 7 to about 14 weeks of age. This was, however, an artefact of data collation. Individual infants showed abrupt falls in heart rate at the time that the adult-like body temperature pattern appeared, but this occurred at different ages in different babies, so when data were collated cross sectionally, an apparently gradual fall resulted. The relation between the developmental changes in sleeping heart rate and rectal temperature was different in boys and girls, with girls showing a more abrupt and greater change in heart rate at the time of development of the adult-like body temperature pattern. Infants whose parents smoked had significantly lower heart rates once the adult-like body temperature pattern had appeared.

Early physiological development of infants with intrauterine growth retardation.

OBJECTIVES: To assess the patterns of early postnatal physiological adaptation and maturation in intrauterine growth retarded (IUGR) infants by measuring changes in sleeping deep body temperature, heart rate, and concentrations of urinary cortisol. SETTING: At home. PATIENTS: Sixty five IUGR babies and 127 controls matched for sex, social class, and levels of parental smoking. RESULTS: Night time sleeping deep body temperature, heart rate, and cortisol excretion fell with age, eventually establishing an adult type diurnal rhythm of physiological function. Minimum overnight temperature showed a linear decline with age (p < 0.001), but the IUGR infants and the controls had significant differences in intercept (p = 0.007) and slope (p = 0.02). The estimated rate of decline per week was 0.020 degrees C for IUGR infants and 0.031 degrees C for controls. Maximum temperature did not show similar changes. IUGR infants had a mean (SE) age adjusted minimum overnight heart rate that was 4.2 (1.5) beats/min (p = 0.005) higher than controls. Overnight cortisol/creatinine ratios declined with age at a rate of 4.1% per week (log ratio -0.421 (0.0165), p = 0.01), but the ratio for IUGR infants was on average 42% higher (log ratio 0.35 (0.11), p = 0.002) than for controls of the same age. Morning cortisol concentrations did not show a similar pattern. CONCLUSIONS: Postnatal physiological adaptation and maturation of IUGR infants is slower than normal and therefore they remain in a physiologically immature state for longer. The higher heart rates and greater cortisol excretion in such infants may be precursors to hypertension and cardiovascular disease seen in adults.

Interactions between infant care practices and physiological development in Asian infants.

Asian infants are less likely to suffer cot death despite apparently higher prevalence of some risk factors. This paper compares the development of night time body temperature patterns in a small sample of Asian babies with the pattern already established for white infants, where babies who develop an adult-like night time temperature pattern later than usual share characteristics with victims of SIDS. The Asian infants had similar body temperature patterns to whites, but tended to develop the adult-like pattern later, not earlier as might have been expected. More Asian infants than white in our sample slept in the parental bed, and, before the adult-like body temperature patterns appeared, co-sleeping infants had higher body temperatures than those in their own cots. Asian infants slept in significantly warmer rooms than whites, but under similar amounts of bedding. These studies do not therefore reveal any physiological difference between Asians and whites which might account for low vulnerability to cot death, indeed, if anything the reverse.

Prone sleeping infants have a reduced ability to lose heat.

The heat loss coefficients of a group of infants have been calculated and compared to see if there is a difference in the ability to lose heat between prone and non-prone sleeping infants. For a group of 43 infants aged 4-29 weeks, a simple mathematical model of exponential cooling in a body has been fitted to the fall in rectal temperature which occurs in infants at bedtime. One of the parameters yielded by the fitting process is the coefficient of thermal heat loss. After validation against the estimated heat loss from supine sleeping infants, the heat loss coefficient was compared at different sleep positions and gender. The mean heat loss coefficient, measured from the non-prone sleeping infants (0.269 W/degrees C, S.D. 0.197) agreed well with the value calculated for supine sleepers with the same tog levels (0.4 W/degrees C). Prone sleeping infants were found to have a considerably smaller heat loss coefficient which was approximately 60% of the value for non-prone sleeping infants (P = 0.000097). Female infants were found to have a heat loss coefficient that was approximately 70% of that of male infants but this gender difference was only significant (P = 0.025) for non-prone sleeping infants. These results suggest that infants sleeping in the prone position may be unable to lose heat as rapidly as those infants sleeping non-prone.

A prospective comparison of computerized arthrotomography and magnetic resonance imaging of the glenohumeral joint.

Twenty-five patients with shoulder instability or shoulder pain of undetermined etiology were prospectively evaluated with magnetic resonance imaging and computerized arthrotomography. Actual lesions were determined by arthroscopy or at the time of open surgical repair. The images obtained were interpreted independently by three radiologists blinded to both surgical results and the results of previous diagnostic tests. Sensitivity, specificity, and accuracy were determined for each imaging technique for a variety of pathologic entities, including anterior and posterior labral abnormalities, capsular redundancy, biceps-labral complex abnormalities, humeral head (Hill-Sachs) impression lesions, and glenohumeral loose bodies. Analysis of imaging techniques also included construction of receiver operator curves for labral abnormalities. Magnetic resonance imaging showed better diagnostic results in the evaluation of glenoid labral and humeral head impression lesions (P < 0.05). Both imaging techniques were equally successful in identifying biceps-labral lesions and intraarticular loose bodies within the glenohumeral joint. Neither imaging technique was consistent in the evaluation of capsular redundancy. Receiver operator curve analysis confirmed that magnetic resonance imaging was the more accurate imaging study in evaluating anterior and posterior glenoid labral abnormalities.

MRI in the evaluation of patients with suspected instability of the shoulder joint including a comparison with CT-arthrography.

Thirty-two patients with clinical suspicion of shoulder instability were the subject of this assessment of MRI (1.5 T G. E. Signa) and its comparison with CT-arthrography (GE8800 and GE9800) of the shoulder. Twenty-seven patients had a final diagnosis established by arthroscopy and five by arthrotomy. The imaging examinations were interpreted independently by three radiologists blinded against history, surgical diagnosis and results of the compared test. The statistical analysis included ROC-curves, sensitivity, specificity and accuracy as well as predictive values. The significance of the comparative results was assessed by the McNemar Sign test. MRI showed significantly better diagnostic results in the evaluation of labral structures (sensitivity 85%, specificity 90%, accuracy 88%) and humeral head impression fractures (75, 82, 78 %). None of the tests were of sufficient diagnostic value to identify capsular abnormalities. CT and MRI results regarding the biceps tendon and loose bodies were similar, however, statistically not significant. MRI proved to be of high diagnostic value in the assessment of the unstable shoulder and compared favorably with CT-A.

Posterior shoulder instability.

Understanding the anatomic restraints to posterior shoulder instability and the resulting pathophysiology helps the treating physician make a correct diagnosis and formulate an appropriate treatment plan. A nonoperative program directed at reducing pain and increasing stability through comprehensive shoulder strengthening methods has generally been successful in treating recurrent posterior shoulder subluxation. Surgical options for treatment are reserved for those patients who fail to recognize improvement after six months of therapy and have no evidence of a psychological disturbance as the cause of their posterior instability.

Revision of failed total shoulder arthroplasty.

The complexity of revision shoulder surgery remains a supreme challenge for the experienced shoulder surgeon. The difficulty of surgery is often accompanied by unpredictable patient cooperation during the postoperative rehabilitation program. Recognition of the problems associated with the failed shoulder arthroplasty is necessary for successful revision surgery. Numerous reconstructive techniques are necessary for restoration of soft tissue and bony deficiencies. Component revision is often necessary in treating component loosening or glenohumeral instability. Glenoid component removal may be necessary in the presence of severe rotator cuff insufficiency or marked glenoid bone deficiency. Humeral revision is most predictably treated with methylmethacrylate fixation. Humeral fractures associated with humeral arthroplasty are most successfully treated surgically, except in those instances where a long oblique or spiral fracture is not associated with prosthetic loosening. Deep infection is most predictably treated by extensive debridement, parenteral antibiotics, and delayed exchange of the components. The success of revision shoulder arthroplasty is often unpredictable, with 60% of revisions offering satisfactory pain relief and restoration of function. Critical to the success of revision arthroplasty is the status of the soft tissues, particularly the anterior deltoid and rotator cuff.

[Pulmonary hypertension in children treated with ventriculo-atrial shunt in hydrocephalus]. / Pulmonal hypertension hos børn behandlet med ventrikulo-atrial shunt for hydrocephalus.

Four infants treated with ventriculo-atrial shunt for hydrocephalus developed sudden pulmonary hypertension, several years after the first operation. All infants died in spite of replacement of the distal shunt and AK treatment. At necropsy, old microemboli were found in the small lung vessels in two infants whereas the two other infants were suspected of having the same lesions from clinical investigations. The overall frequency for this complication was estimated to be 6.7% (95% CI 1.4-14.3%).