Erratum: Measurement of Body-Centered-Cubic Aluminum at 475 GPa [Phys. Rev. Lett. 119, 175702 (2017)].

This corrects the article DOI: 10.1103/PhysRevLett.119.175702.

Measurement of Body-Centered-Cubic Aluminum at 475 GPa.

Nanosecond in situ x-ray diffraction and simultaneous velocimetry measurements were used to determine the crystal structure and pressure, respectively, of ramp-compressed aluminum at stress states between 111 and 475 GPa. The solid-solid Al phase transformations, fcc-hcp and hcp-bcc, are observed at 216±9 and 321±12 GPa, respectively, with the bcc phase persisting to 475 GPa. The high-pressure crystallographic texture of the hcp and bcc phases suggests close-packed or nearly close-packed lattice planes remain parallel through both transformations.

Morbidity after adenotonsillectomy for paediatric obstructive sleep apnoea syndrome: waking up to a pragmatic approach.

INTRODUCTION: Adenotonsillectomy is successful at eliminating airway obstruction in the majority of otherwise normal children with obstructive sleep apnoea syndrome. Children with this condition are at significantly higher risk of post-operative respiratory complications. Identifying children at risk of post-operative respiratory complications after adenotonsillectomy for obstructive sleep apnoea syndrome remains a challenge for clinicians, especially those at district general hospitals. AIM: To review the evidence and to proffer a pragmatic approach to diagnosis and management, by classifying those at risk of post-operative respiratory complications into different risk subsets, with guidelines for management. CONCLUSION: Patients in the high risk group should be operated upon at paediatric specialist centres with intensive care facilities. Those in the moderate risk group may undergo adenotonsillectomy at their district general hospital, provided facilities for administering continuous positive airway pressure are available on-site. Most children with obstructive sleep apnoea syndrome may be classified as low risk candidates and may safely be operated upon at their local district general hospital.

Effects of KHEYLRFamide and KNEFIRFamide on cyclic adenosine monophosphate levels in Ascaris suum somatic muscle.

KHEYLRF-NH(2) (AF2) is a FMRFamide-related peptide (FaRP) present in parasitic and free-living nematodes. At concentrations as low as 10 pM, AF2 induces a biphasic tension response, consisting of a transient relaxation followed by profound excitation, in neuromuscular strips prepared from Ascaris suum. In the present study, the effects of AF2 on cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP) and inositol-1,4,5-triphosphate (IP(3)) levels were measured following muscle tension recordings from 2 cm neuromuscular strips prepared from adult A. suum. AF2 induced a concentration- and time-dependent increase in cAMP, beginning at 1 nM; cAMP levels increased by 84-fold following 1 h exposure to 1 microM AF2. cGMP and IP(3) levels were unaffected by AF2 at concentrations

Laser Doppler flux-metry in laryngeal squamous cell carcinoma.

Tumour angiogenesis has recently attracted a great deal of attention as a critical part of oncogenesis and a necessary prerequisite for a malignant phenotype. Novel antiangiogenic therapy for solid tumours including laryngeal cancer is entering clinical trials. Quantifying microvessel density is considered the gold standard for measuring baseline angiogenesis and indeed 'the response to intervention'. We hypothesize that laser Doppler flux-metry could provide a non-invasive reliable method of quantifying blood flux within tumours. The aims were to determine whether a laser Doppler flux meter could be used as a reliable and reproducible method of estimating blood flux in the human larynx and to establish baseline Doppler flux recordings for the human larynx. The method used was a validation study in patients with laryngeal squamous cell cancer and normal controls. Statistical analysis was performed using SPSS software. We have demonstrated good reproducibility of laser Doppler measurements in human laryngeal mucosa (correlation coefficient 0.956 @P = 0.01). We have also derived arbitrary means of laser Doppler flux-metry in normal laryngeal mucosa and in squamous cell carcinoma of the larynx. Comparisons between normal and tumour laser Doppler flux-metry (LDF) readings showed no significant difference. We suggest that Laser Doppler flux-metry is a potentially useful tool with which to study blood flow in the larynx and propose arbitrary LDF levels for the normal and diseased human larynx.

Burkholderia cepacia lower respiratory tract infection associated with exposure to a respiratory therapist.

OBJECTIVE: To investigate and control a nosocomial outbreak of Burkholderia cepacia lower respiratory tract infection. DESIGN: Outbreak investigation and case-control study. SETTING: A 260-bed community hospital. PATIENTS: Participants were mechanically ventilated intensive care patients without cystic fibrosis. A case was defined as a hospitalized patient with a sputum culture positive for B. cepacia between January 1 and November 6, 1998. METHODS: Respiratory therapy infection control policies and practices were reviewed; laboratory and environmental studies and a retrospective case-control study were conducted. Case-patients were matched with control-patients on age, gender, diagnosis, and type of intensive care unit. RESULTS: Nine case-patients were identified; B. cepacia likely caused pneumonia in seven and colonization in two. Two respiratory therapy practices probably contributed to the transmission of B. cepacia: multidose albuterol vials were used among several patients, and nebulizer assemblies often were not dried between uses. B. cepacia was grown from cultures of three previously opened multidose vials; pulsed-field gel electrophoresis patterns of B. cepacia from seven case-patients and two multidose vials were indistinguishable. Case-patients had longer durations of heated humidified mechanical ventilation (mean, 9.8 days vs 4.4 days; P=.03) and were more likely to have exposure to one particular respiratory therapist than controls (odds ratio, undefined; 95% confidence interval, 4.7-infinity; P=.001). The association with the respiratory therapist, a temporary employee, persisted after controlling for duration of heated humidified ventilation. No new B. cepacia infections were identified after control measures were implemented. CONCLUSIONS: B. cepacia probably was transmitted among patients through use of extrinsically contaminated multidose albuterol vials. Respiratory therapy departments must pay close attention to infection control practices, particularly among new or temporary staff.

Understanding screening: requirements for a successful programme.

Screening has generally been successful in identifying those at risk from disease. This success has led to the belief that screening in the general population is always a good thing. However, there are pitfalls which must be avoided if screening programmes are to achieve what is intended for them.

Juvenile xanthogranuloma of the nasal cavity.

Juvenile xanthogranuloma is a benign, normolipaemic, self-healing condition and a type of histiocytosis that occurs most frequently in infants and children. This condition usually presents with solitary or multiple cutaneous lesions and occasionally with visceral lesions. We report a case of juvenile xanthogranuloma occurring in the nasal cavity. We believe this is the first report, in the English literature, of juvenile xanthogranuloma occurring in this site.

Characterizing the response of calcium signal transducers to generated calcium transients.

Cellular Ca2+ transients and Ca2+-binding proteins regulate physiological phenomena as diverse as muscle contraction, neurosecretion, and cell division. When Ca2+ is rapidly mixed with slow Ca2+ chelators, EGTA, or Mg2+/EDTA, artificial Ca2+ transients (ACTs) of varying duration (0.1-50 ms half-widths (hws)) and amplitude can be generated. We have exposed several Ca2+ indicators, Ca2+-binding proteins, and a Ca2+-dependent enzyme to ACTs of various durations and observed their transient binding of Ca2+, complex formation, and/or activation. A 0.1 ms hw ACT transiently occupied approximately 70% of the N-terminal regulatory sites of troponin C consistent with their rapid Ca2+ on-rate (8.7 +/- 2.0 x 10(7) M-1 s-1). A 1.1 ms hw ACT produced approximately 90% transient binding of the N-terminal of calmodulin (CaM) to the RS-20 peptide, but little binding of CaM's C-terminal to RS-20. A 0.6 ms hw ACT was sufficient for the N-terminal of CaM to transiently bind approximately 60% of myosin light chain kinase (MLCK), while a 1.8 ms hw ACT produced approximately 22% transient activation of the sarcoplasmic reticulum (SR) Ca2+/ATPase. In both cases, the ACT had fallen back to baseline approximately 10-30 ms before maximal binding of CaM to MLCK or SR Ca2+/ATPase activation occurred and binding and enzyme activation persisted long after the Ca transient had subsided. The use of ACTs has allowed us to visualize how the Ca2+-exchange rates of Ca2+-binding proteins dictate their Ca2+-induced conformational changes, Ca2+-induced protein/peptide and protein/protein interactions, and enzyme activation and inactivation, in response to Ca2+ transients of various amplitude and duration. By characterizing the response of these proteins to ACTs, we can predict with greater certainty how they would respond to natural Ca2+ transients to regulate cellular phenomena.

Comparative in vitro effects of closantel and selected beta-ketoamide anthelmintics on a gastrointestinal nematode and vertebrate liver cells.

PNU-87407 and PNU-88509, beta-ketoamide anthelmintics that are structurally related to each other and to the salicylanilide anthelmintic closantel, exhibit different anthelmintic spectra and apparent toxicity in mammals. The basis for this differential pharmacology was examined in experiments that measured motility and adenosine triphosphate (ATP) levels in larval and adult stages of the gastrointestinal nematode, Haemonchus contortus, and in a vertebrate liver cell line and mitochondria. PNU-87407 and PNU-88509 both exhibited functional cross-resistance with closantel in larval migration assays using closantel-resistant and -sensitive isolates of H. contortus. Each compound reduced motility and ATP levels in cultured adult H. contortus in a concentration- and time-dependent manner; however, motility was reduced more rapidly by PNU-88509, and ATP levels were reduced by lower concentrations of closantel than the beta-ketoamides. Tension recordings from segments of adult H. contortus showed that PNU-88509 induces spastic paralysis, while PNU-87407 and closantel induce flaccid paralysis of the somatic musculature. Marked differences in the actions of these compounds were also observed in the mammalian preparations. In Chang liver cells, ATP levels were reduced after 3 h exposures to > or = 0.25 microM PNU-87407, > or = 1 microM closantel or > or = 10 microM PNU-88509. Reductions in ATP caused by PNU-88509 were completely reversible, while the effects of closantel and PNU-87407 were irreversible. PNU-87407, closantel and PNU-88509 uncoupled oxidative phosphorylation in isolated rat liver mitochondria, inhibiting the respiratory control index (with glutamate or succinate as substrate) by 50% at concentrations of 0.14, 0.9 and 7.6 microM, respectively.

Histidine to alanine mutants of human dihydroorotate dehydrogenase. Identification of a brequinar-resistant mutant enzyme.

Dihydroorotate dehydrogenase (DHODase) is the rate-limiting enzyme of the mammalian de novo pyrimidine biosynthesis pathway, and is the molecular target of the antiproliferative, immunosuppressive compound brequinar sodium (BQR). We have shown previously that the activity of the recombinant human enzyme displays pH and diethylpyrocarbonate sensitivities that implicate a critical role for one or more histidine residues in catalysis [Copeland et al., Arch Biochem Biophys 323: 79-86, 1995.]. Here we report the results of alanine scanning mutagenesis for each of the 8 histidine residues of the recombinant human enzyme. In most cases, the replacement of histidine by alanine had little effect on the Km values of the two substrates, dihydroorotate and ubiquinone, or on the overall kcat of the enzymatic reaction. Replacement of H71, H129, and H364 by alanine, however, completely abolished enzymatic activity. The loss of activity for the H71A mutant was unexpected, since this residue is not conserved in the homologous rat enzyme; in the rodent enzyme this residue is an asparagine. Replacement of H71 by asparagine in the human enzyme led to a full recovery of enzymatic activity, indicating that a histidine is not required at this position. Replacement of H26 by alanine led to about a 10-fold reduction in catalytic activity relative to the wild-type enzyme, with no significant perturbation of the substrate Km values. This mutant was, however, at least 167-fold less sensitive to inhibition by the noncompetitive inhibitor BQR. While the wild-type and other mutant enzymes displayed IC50 values for BQR inhibition between 6 and 10 nM, the H26A mutant was inhibited less than 25% at concentrations of BQR as high as 150 nM. These data suggest that H26 plays an important role in BQR binding to the enzyme.

Alterations in inotropy, nitric oxide and cyclic GMP synthesis, protein phosphorylation and ADP-ribosylation in the endotoxin-treated rat myocardium and cardiomyocytes.

To evaluate the effects of the in vivo endotoxin treatment of the rat on (1) the contractile responses in the subsequently isolated papillary muscle to adrenergic and cholinergic agonists and (2) the biochemical parameters (cyclic GMP, nitric oxide synthesis, protein phosphorylation and ADP-ribosyslation) in the subsequently isolated cardiomyocytes. Following the in vivo endotoxin treatment (4 mg/kg i.p., 18 h), contractile responses to increasing amounts of isoprenaline or to increasing amounts of oxotremorine in the presence of a fixed amount of isoprenaline were determined in isolated papillary strips. Activities of nitric oxide synthase, guanylyl cyclase, as well as phosphorylation of phospholamban and troponin-inhibitory subunit, and pertussis toxin-catalyzed and endogenous ADP-ribosylations were determined in the intact cardiomyocytes and subcellular fractions. The increase in the force of contraction by isoprenaline was reduced, while its inhibition by oxotremorine was greater in the endotoxin-treated papillary strips. The activities of both nitric oxide synthase, primarily of the inducible form of the enzyme, and cytosolic guanylyl cyclase were higher while the phosphorylations of both phospholamban and troponin-inhibitory subunit were of lesser magnitude in the cardiomyocytes following the in vivo endotoxin treatment. Pertussis toxin-catalyzed ADP-ribosylation of the 41 kDa polypeptide, which is the alpha subunit of Gi, was also decreased. The results of the present study support the postulate that alterations in both the cyclic AMP and cyclic GMP signalling cascade contribute to the myocardial dysfunction caused by endotoxin and cytokines.

Anthelmintic profile of the cyclodepsipeptide PF1022A in in vitro and in vivo models.

A novel cyclodepsipeptide of fungal origin, PF1022A, recently was reported to have anthelmintic activity. To supplement published reports and determine potential utility of PF1022A as a ruminant anthelmintic, the compound was examined in in vitro and in vivo models. Assays used measured motility of Haemonchus contortus (intrinsic drug potency), ATP levels (parasite death), and activity against H. contortus, Ostertagia ostertagi, and Trichostrongylus colubriformis in the jird (spectrum, potency, and efficacy by various routes). The potency of PF1022A in reducing motility is greater than commercial anthelmintics. Examination of ATP levels in PF1022A-paralyzed H. contortus indicates that worms are not killed, suggesting the compound acts as a neurotoxin in nematodes. In the jird, PF1022A has activity orally against each of the parasites studied and at doses comparable to all commercial anthelmintics, except the macrocyclic lactones which are more potent. Unfortunately, for some nematode species, parenteral delivery is ineffective at realistic doses.

The hyoid syndrome: a pain in the neck.

This paper reports on 13 patients with a pain syndrome arising from the region of the greater cornu of the hyoid bone. It is often missed and yet is readily treatable. The syndrome causes pain on swallowing in the region of the hyoid which may radiate to the ear, face and lower jaw or may be felt also in the pharynx. Treatment by an injection of a mixture of depomedrone and one per cent lignocaine is very effective. The underlying pathology is discussed and it is suggested that in some cases the pain and discomfort experienced by patients may be due to tenosynovitis of the intermediate tendon of the digastric muscle. Greater recognition of this relatively common condition would not only result in more effective treatment but would also avoid unnecessary investigation and surgery.

Oropharyngeal trauma during routine ear, nose and throat procedures.

A prospective study of 122 adults undergoing routine ear, nose and throat (ENT) operations over a three-month period was carried out to determine the cause of oropharyngeal trauma seen in some patients. Those having tonsillar or palatal surgery were excluded from the study. Forty-five (36.9 per cent) patients complained of a mild sore throat post-operatively and six (4.9 per cent) of a severe sore throat. Five of these (4 per cent) had evidence of injury to the uvula and soft palate which delayed their discharge from hospital. No single cause of trauma was identified but possible factors included the use of laryngeal masks (two cases), throat packs (two cases) and blind suctioning with a plastic Yankauer sucker (one case). There is a relatively high risk of oropharyngeal trauma during routine otolaryngological procedures and we recommend that care should be taken to prevent this common cause of significant post-operative morbidity.

Kinetic and site-directed mutagenesis studies of the cysteine residues of bovine low molecular weight phosphotyrosyl protein phosphatase.

The roles of the 8 conserved cysteines and 1 arginine in the low molecular weight phosphotyrosyl protein phosphatases were investigated using site-directed mutagenesis of the recombinant bovine heart enzyme. Single mutants of cysteine to serine were studied for each cysteine; alanine replacements were also made for Cys-12, Cys-17, and Arg-18. The CD spectra of the purified proteins were effectively superimposable, consistent with the conclusion that no major structural alterations had occurred, but 1H NMR spectroscopy did reveal some spectral shifts in the aromatic region. Kinetic analysis of the mutant proteins demonstrated that only Cys-12, Cys-17, and Arg-18 had significantly altered catalytic activity toward the substrate p-nitrophenyl phosphate at pH 5. The Cys-12 and Arg-18 mutants were effectively inactive. Thus, it is concluded that Cys-12 is the catalytic nucleophile, and Arg-18 presumably serves an essential function in substrate binding. The C17S mutant had 6% residual activity compared with wild type protein, whereas the C17A mutant had 37% activity. Consistent with the observed activity of the Cys-17 mutant, a covalent phosphocysteine intermediate was trapped and identified by 31P NMR. Further kinetic analysis of C17A using several aryl phosphate monoester substrates with different leaving group pK alpha values indicated that no change in the rate-determining step of the catalytic mechanism had occurred, that is, dephosphorylation of the covalent phosphoenzyme intermediate remains rate-limiting. The C17A mutant had a 4-fold higher phosphate Ki and slightly higher Km values for p-nitrophenyl phosphate suggesting that Cys-17 may be important for optimal positioning of the substrate phosphate moiety.