Salt marsh vegetation change during a half-century of experimental nutrient addition and climate-driven controls in Great Sippewissett Marsh.

Vegetative cover was mapped annually, 1976-2022, in experimental plots in Great Sippewissett Marsh, Cape Cod, USA, chronically fertilized at different doses, and subject to changes in sea level and other climate-related variables. Dominant species within areas of higher elevation in the plots followed different decadal trajectories: rise in sea level diminished cover of Spartina patens; higher N supplies increased cover of Distichlis spicata. The opportunistic growth response of D. spicata to high N supplies unexpectedly fostered increased platform accretion, a feature that persisted for succeeding decades and led to further changes in vegetation: D. spicata functioned as an effective ecosystem engineer with long-term ecological consequences. Shrubs usually found in upper marsh margins expanded into areas where D. spicata had stimulated accretion, then shaded and excluded D. spicata, but subsequently lost cover as sea level rise continued. Increased N supply converted stands of Spartina alterniflora, the dominant low marsh species, from short to taller ecophenotypes; sea level rise had minor effects on S. alterniflora, but during 2019-2022 appeared to reach a tipping point that fostered taller S. alterniflora and bare space even in un-fertilized control plots, and in Great Sippewissett Marsh in general. Model results anticipate that-in spite of potential accretion enhanced by vegetation and ecosystem engineer effects-there will be loss of high marsh, transient increases of low marsh, followed by loss of low marsh, and eventual conversion to shallow open water by the end of the century. Dire local projections match those of the plurality of recent reports from salt marshes around the world. Proposed management strategies may only delay unfortunate outcomes rather than maintain wetlands. Concerted reductions of warming from greenhouse gases, and lower N loads seem necessary to address the coming crises in wetlands-and many other environmental threats.

LMA Supreme insertion by novices in manikins and patients.

The LMA Supreme has been suggested for use in emergency situations by medical personnel with no experience in endotracheal intubation. We evaluated the LMA Supreme when inserted by non-anaesthetists, firstly in a manikin and then in patients. Fifty airway novices inserted a LMA Supreme in a manikin without any complications so we proceeded to the patient phase. Fifty airway novices inserted the LMA Supreme in anaesthetised patients undergoing elective surgery. First time insertion success rate was 86% and overall insertion success rate was 100%. Mechanical ventilation was successful in all cases. Median (IQR [range]) time to establish an airway was 34 s (26-40 [18-145] s). Median (IQR [range]) pharyngeal seal pressure was 23 cmH(2)O (19-28 [13-40] cmH(2)O). There were no important complications. Results are consistent with previous studies of use of the LMA Supreme by airway experts. We conclude that the LMA supreme is suitable for use by airway novices. Further research is needed before it may be recommended for cardiopulmonary resuscitation and emergency airway use.

A low-cost, endoscopic, digital, still and video photography system for ENT clinics.

Image capture systems that display and record endoscopic images are important for documentation and teaching. We have modified a universal serial bus microscope to couple with most clinical endoscopes used in our practice. This very economical device produces images suitable for teaching, and potentially for clinical use. The implications of this could be significant for teaching, patient education, documentation and the developing world.

The reliability of laryngoscope lights.

The aim of this study is to compare the brightness of laryngoscope light between instruments, and the variability within the same instrument. A secondary aim is to compare the light from a reusable laryngoscope with a single use design. Testing equipment was constructed which provided a qualitative measure of light output from laryngoscopes. Anaesthetists assessed the minimum acceptable brightness for intubation using the intubating mannikin and a variable brightness laryngoscope. Laryngoscopes in anaesthetic rooms around the hospital were then tested repeatedly, and the results were compared to the anaesthetists' opinions. There was considerable intersubject and intrasubject variation in the reusable design, often failing to satisfy the minimum brightness. The single use blades were found to be brighter and more reliable than reusable blades.

Mutation of residues critical for benzohydroxamic acid binding to horseradish peroxidase isoenzyme C.

Aromatic substrate binding to peroxidases is mediated through hydrophobic and hydrogen bonding interactions between residues on the distal side of the heme and the substrate molecule. The effects of perturbing these interactions are investigated by an electronic absorption and resonance Raman study of benzohydroxamic acid (BHA) binding to a series of mutants of horseradish peroxidase isoenzyme C (HRPC). In particular, the Phe179 --> Ala, His42 --> Glu variants and the double mutant His42 --> Glu:Arg38 --> Leu are studied in their ferric state at pH 7 with and without BHA. A comparison of the data with those previously reported for wild-type HRPC and other distal site mutants reaffirms that in the resting state mutation of His42 leads to an increase of 6-coordinate aquo heme forms at the expense of the 5-coordinate heme state, which is the dominant species in wild-type HRPC. The His42Glu:Arg38Leu double mutant displays an enhanced proportion of the pentacoordinate heme state, similar to the single Arg38Leu mutant. The heme spin states are insensitive to mutation of the Phe179 residue. The BHA complexes of all mutants are found to have a greater amount of unbound form compared to the wild-type HRPC complex. It is apparent from the spectral changes induced on complexation with BHA that, although Phe179 provides an important hydrophobic interaction with BHA, the hydrogen bonds formed between His42 and, in particular, Arg38 and BHA assume a more critical role in the binding of BHA to the resting state.

The critical role of the proximal calcium ion in the structural properties of horseradish peroxidase.

The extent to which the structural Ca(2+) ions of horseradish peroxidase (HRPC) are a determinant in defining the heme pocket architecture is investigated by electronic absorption and resonance Raman spectroscopy upon removal of one Ca(2+) ion. The Fe(III) heme states are modified upon Ca(2+) depletion, with an uncommon quantum mechanically mixed spin state becoming the dominant species. Ca(2+)-depleted HRPC forms complexes with benzohydroxamic acid and CO which display spectra very similar to those of native HRPC, indicating that any changes to the distal cavity structural properties upon Ca(2+) depletion are easily reversed. Contrary to the native protein, the Ca(2+)-depleted ferrous form displays a low-spin bis-histidyl heme state and a small proportion of high-spin heme. Furthermore, the nu(Fe-Im) stretching mode downshifts 27 cm(-1) upon Ca(2+) depletion revealing a significant structural perturbation of the proximal cavity near the histidine ligand. The specific activity of the Ca(2+)-depleted enzyme is 50% that of the native form. The effects on enzyme activity and spectral features observed upon Ca(2+) depletion are reversible upon reconstitution. Evaluation of the present and previous data firmly favors the proximal Ca(2+) ion as that which is lost upon Ca(2+) depletion and which likely plays the more critical role in regulating the heme pocket structural and catalytic properties.

Insulin-like growth factor-I augments prolactin and inhibits growth hormone release through distinct as well as overlapping cellular signaling pathways.

We recently discovered a new role for insulin-like growth factor-I (IGF-I) as a specific and direct stimulator of prolactin (PRL) release in addition to its recognized function as an inhibitor of growth hormone (GH) release and synthesis. Little is known of the mechanisms that transduce the actions of IGF-I on PRL and GH release in vertebrates. The present study was undertaken to determine the cellular pathways that mediate the disparate actions of IGF-I on PRL and GH release in hybrid striped bass (Morone saxatilis X M. chrysops). When regulating cellular function, IGF-I may activate two primary pathways, phosphatidylinositol 3-kinase (PI 3-K) and mitogen-activated protein kinase (MAPK). The specific MAPK inhibitor, PD98059, blocked IGF-I-evoked PRL release as well as GH release inhibition over an 18-20-h incubation. LY294002, a specific PI 3-K inhibitor, overcame IGF-I's inhibition of GH release but was ineffective in blocking PRL release stimulated by IGF-I. These studies suggest IGF-I disparately alters PRL and GH by activating distinct as well as overlapping signaling pathways central for mediating actions of growth factors on secretory activity as well as cell proliferation. These results further support a role for IGF-I as a physiological regulator of PRL and GH.

Haem-linked interactions in horseradish peroxidase revealed by spectroscopic analysis of the Phe-221-->Met mutant.

A gene encoding a Phe-221-to-Met substitution in the haem enzyme horseradish peroxidase has been constructed and expressed in Escherichia coli. In the wild-type enzyme the side chain of Phe-221 is tightly stacked against the imidazole ring of His-170, which provides the only axial ligand to the haem iron atom. The Phe-221-->Met enzyme is active, and forms characteristic complexes with typical peroxidase ligands (CO, cyanide, fluoride), and with benzhydroxamic acid. Significant differences between the mutant and wild-type enzymes can be detected spectroscopically. These include a change in the Fe(III) resting state of the enzyme to an unusual quantum mechanically mixed-spin haem species, a marked decrease in the pK(a) of the alkaline transition and a reduction in enzyme stability at alkaline pH for both Fe(III) and Fe(II) forms. The perturbation of the haem pocket in the mutant can be attributed to several factors, including the increased steric freedom and solvent accessibility of the His-170 ligand, as indicated by (1)H-NMR data, and the loss of the pi-pi interaction between His-170 and Phe-221.

Effect of low temperature on soybean peroxidase: spectroscopic characterization of the quantum-mechanically admixed spin state.

A spectroscopic study of soybean peroxidase (SBP) has been carried out using electronic absorption, resonance Raman (RR) and electron paramagnetic resonance (EPR) spectroscopy in order to determine the effects of temperature on the heme spin state. Upon lowering the temperature a transition from high spin to low spin is induced in SBP resulting from conformational changes in the heme cavity, including a contraction of the heme core, the reorientation of the vinyl group in position 2 of the porphyrin macrocycle, and the binding of the distal His to the Fe atom. Moreover, the combined analysis of the data derived from the different techniques at both room and low temperatures demonstrates that at low temperature the quantum-mechanically admixed spin state (QS) of SBP has RR frequencies different from those observed for the QS species at room temperature.

Formation of two types of low-spin heme in horseradish peroxidase isoenzyme A2 at low temperature.

Electronic absorption, resonance Raman and EPR spectra are reported for ferric horseradish peroxidase isoenzyme A2 at neutral and alkaline pH together with its imidazole complex at 12 K. The data are compared with those obtained at room temperature. At neutral pH, lowering the temperature induces conformational changes with the formation of two types of low-spin hemes, a bis-histidyl type and a hydroxo type. The transition induced by lowering the temperature is accompanied by a change in the orientation of a vinyl substituent which appears less conjugated to the porphyrin macrocycle than at room temperature. At low temperature the low-spin hemes coexist with a quantum admixed spin species. All the forms are characterized by extremely high resonance Raman frequencies, indicating a contraction of the core size from that of the room temperature species. At alkaline pH, only one low-spin species is observed at both room and low temperatures, with a hydroxo ligand bound to the heme iron. The v(Fe-OH) stretching mode has been assigned at 512 cm(-1), on the basis of the isotopic shift observed in D2O and H2(18)O. This relatively low frequency, together with the anomalous shift observed in deuterium, indicates that the hydrogen bonds between the oxygen atom and the distal residues are stronger than in metmyoglobin, but weaker than those of horseradish peroxidase isoenzyme C. This is in agreement with the lower tetragonality, determined from the EPR g values, of alkaline horseradish peroxidase isoenzyme A2 than of metmyoglobin.

Treatment of low back pain associated with ;;back mice'': a case series.

Back mice are subcutaneous fibroadenomatous nodules that cause low back symptoms. Previous case reports do not provide systematic descriptions of the clinical presentation or long-term follow-up of this problem. This retrospective case series reports syndrome characteristics and treatment outcomes for injection therapy for "back mice." We completed telephone interviews, chart reviews, and written questionnaires for a convenience sample of 35 participants. Participants reported the following symptoms: pain radiating to the lower leg (37%), leg numbness or paresthesias (14%), and a median of 8 weeks of pain before treatment (range 3 weeks to 10 years). Thirty-one participants (89%) received lasting relief from injection of local anesthetic and corticosteroid. Injection therapy relieved both local and radiating symptoms but often did not eliminate the nodules. Thirty participants (86%) were "satisfied" or "very satisfied" with the treatment. There were no adverse events reported. Back mice can cause radiating pain that can be confused with other low back or leg syndromes. Injection treatment seems to be effective, long lasting, and well tolerated. Physicians should search for these nodules in patients with unexplained low back pain and try injection therapy before initiating expensive therapy.

The quantum mixed-spin heme state of barley peroxidase: A paradigm for class III peroxidases.

Electronic absorption and resonance Raman (RR) spectra of the ferric form of barley grain peroxidase (BP 1) at various pH values, at both room temperature and 20 K, are reported, together with electron paramagnetic resonance spectra at 10 K. The ferrous forms and the ferric complex with fluoride have also been studied. A quantum mechanically mixed-spin (QS) state has been identified. The QS heme species coexists with 6- and 5-cHS hemes; the relative populations of these three spin states are found to be dependent on pH and temperature. However, the QS species remains in all cases the dominant heme spin species. Barley peroxidase appears to be further characterized by a splitting of the two vinyl stretching modes, indicating that the vinyl groups are differently conjugated with the porphyrin. An analysis of the currently available spectroscopic data for proteins from all three peroxidase classes suggests that the simultaneous occurrence of the QS heme state as well as the splitting of the two vinyl stretching modes is confined to class III enzymes. The former point is discussed in terms of the possible influences of heme deformations on heme spin state. It is found that moderate saddling alone is probably not enough to cause the QS state, although some saddling may be necessary for the QS state.

pH-dependence for binding a single nitrite ion to each type-2 copper centre in the copper-containing nitrite reductase of Alcaligenes xylosoxidans.

The first quantitative characterization of the interaction of NO2(-) with the Cu-containing dissimilatory nitrite reductase (NiR) of Alcaligenes xylosoxidans using steady-state kinetics, equilibrium gel filtration and EPR spectroscopy is described. Each molecule of this protein consists of three equivalent subunits, each containing a type-1 Cu atom and also a type-2 Cu atom at each subunit interface. Enzyme activity increased in a biphasic manner with decreasing pH, having an optimum at pH 5.2 and a plateau between pH 6.1 and 5.8. Equilibrium gel filtration showed that binding of NO2(-) to the oxidized NiR was also pH-dependent. At pH 7.5, no binding was detectable, but binding was detectable at lower pH values. At pH 5.2, the concentration-dependence for binding of NO2(-) to the enzyme showed that approx. 4.1 NO2(-) ions bound per trimeric NiR molecule. Unexpectedly, NiR deficient in type-2 Cu centres bound 1.3 NO2(-) ions per trimer. When corrected for this binding, a value of 3 NO2(-) ions bound per trimer of NiR, equivalent to the type-2 Cu content. The NO2(-)-induced changes in the EPR parameters of the type-2 Cu centre of the oxidized enzyme showed a similar pH-dependence to that of the activity. Binding constants for NO2(-) at a single type of site, after allowing for the non-specifically bound NO2(-), were 350+/-35 microM (mean+/-S.E.M.) at pH 7.5 and <30 microM at pH 5.2. The apparent Km for NO2(-) with saturating concentrations of dithionite as reductant was 35 microM at pH 7.5, which is 10-fold tighter than for the oxidized enzyme, and is compatible with an ordered mechanism in which the enzyme is reduced before NO2(-) binds.