Inclusion of Trainee Stakeholders Is Necessary for Effective Change in Health-Service-Psychology Internship Training.

In a recent call to action, we described pressing issues in the health-service-psychology (HSP) internship from the perspective of interns. In our article, we sought to initiate a dialogue that would include trainees and bring about concrete changes. The commentaries on our article are a testament to the readiness of the field to engage in such a dialogue, and we applaud the actionable recommendations that they make. In our response to these commentaries, we seek to move the conversation further forward. We observe two themes that cut across these responses: the impetus to gather novel data on training (the "need to know") and the importance of taking action (the "need to act"). We emphasize that in new efforts to gather data and take policy-level action, the inclusion of trainee stakeholders (as well as others involved in and affected by HSP training) is a crucial ingredient for sustainable and equitable change.

Systemic Challenges in Internship Training for Health-Service Psychology: A Call to Action From Trainee Stakeholders.

The challenges observed in health service psychology (HSP) training during COVID-19 revealed systemic and philosophical issues that preexisted the pandemic, but became more visible during the global health crisis. In a position paper written by 23 trainees across different sites and training specializations, the authors use lessons learned from COVID-19 as a touchstone for a call to action in HSP training. Historically, trainee voices have been conspicuously absent from literature about clinical training. We describe longstanding dilemmas in HSP training that were exacerbated by the pandemic and will continue to require resolution after the pandemic has subsided. The authors make recommendations for systems-level changes that would advance equity and sustainability in HSP training. This article advances the conversation about HSP training by including the perspective of trainees as essential stakeholders.

Short communication: Effect of a stable pen management strategy for precalving cows on dry matter intake, plasma nonesterified fatty acid levels, and milk production.

During the close-up transition period, dairy cows are at risk for negative energy balance due to increasing energy demands and decreasing feed intake. This can result in postparturient health problems and decreased milk production after calving. Cows are frequently regrouped during this period, which can negatively affect feeding and resting behavior. The hypothesis was that housing in a stable pen during the close-up transition period should result in a more settled environment resulting in fewer displacements from the feed bunk, which would result in improved feed intake, energy balance [lower nonesterified fatty acid (NEFA) concentrations], and lactation performance. This study addresses precalving pen grouping strategies, which have the potential to affect feed intake and energy balance. A randomized complete block design with pen as the experimental unit was used to compare a stable (S) housing strategy (cows with similar calving dates added to a precalving pen at once) to the more traditional dynamic (D) housing strategy (cows added up to 2 times per week to a precalving pen). Twice-weekly blood samples were collected for NEFA analysis and cow interactions within the pen were observed. Dry matter intake (DMI), milk production, and postparturient health problems were recorded. Mean DMI for the duration of the 28 d of the study was not different (S: 25.5 ± 1.6 vs. D: 25.7 ± 1.0 kg/d), and when examined over time relative to calving, no treatment by time interaction was observed. Concentrations of NEFA were not different when cows initially entered the pens (S: 0.21 ± 0.10 vs. D: 0.18 ± 0.04 mEq/L) and remained not different for the time intervals closer to calving (d -9 to -14: S: 0.28 ± 0.09 vs. D: 0.21 ± 0.04; d -3 to -6: S 0.36 ± 0.10, D 0.32 ± 0.05 mEq/L). Pen grouping strategy did not affect DMI, plasma NEFA concentrations, or milk production.

Structure/function relationships in nickel metallobiochemistry.

Among the many highlights of nickel metallobiochemistry in 1998 were the discoveries that Escherichia coli glyoxalase I is the first example of a nickel isomerase, and that the superoxide dismutase isolated from Streptomyces seoulensis is a new structural class of superoxide dismutase that features thiolate ligation.

An in vitro biomechanical evaluation of bone cements used in percutaneous vertebroplasty.

The purpose of this study was to determine the strength and stiffness of osteoporotic vertebral bodies (VBs) subjected to compression fractures and subsequently treated with bipedicular injections of various polymethylmethacrylate cements. Ten spines were harvested from nonembalmed female cadavers (age 68.6 +/- 13.7 years) and evaluated for bone mineral density using the dual energy X-ray absorptiometry method (t-score = -2.3 +/- 2.4). The 50 VBs (L1-L5) were disarticulated, compressed in a materials testing machine to determine initial strength and stiffness, and then assigned to one of six groups. Two of these groups (n = 8, n = 9) concerned experimental cements, the results of which are not reported here. The 33 vertebral bodies in the remaining four groups were left untreated or were repaired using a transpedicular injection of one of three commercially available polymethylmethacrylate cements. These four groups were: a) no treatment (no cement, n = 8); b) Simplex P (n = 9); c) Cranioplastic (n = 8); and d) Osteobond (n = 8). The VBs were then compressed again according to the initial protocol, and posttreatment strength and stiffness were measured. Results suggested that bipedicular injection of Simplex P and Osteobond restored VB stiffness to initial values, whereas VBs injected with Cranioplastic were significantly less stiff than in their initial state. VBs injected with cement (regardless of type) were significantly stronger than they were initially.

A randomized and blinded field trial to assess the efficacy of an autogenous vaccine to prevent naturally occurring infectious bovine keratoconjunctivis (IBK) in beef calves.

A randomized and blinded 2-arm parallel trial was conducted to assess the efficacy of an autogenous vaccine to prevent naturally occurring infectious bovine keratoconjunctivis (IBK) in beef calves. The trial was managed between May and November 2008 on university owned farms in Iowa and Wisconsin. The vaccine at Iowa contained Moraxella bovoculi (M. bovoculi) while the organism used in the Wisconsin herds vaccine was Branhemella ovis (B. ovis renamed M. ovis). Calves born between January and May 2008 without visible corneal lesions were randomized to receive an autogenous vaccine or placebo vaccine using a computer generated sequence. Two subcutaneous doses were administered 21-28 days apart. Allocation to treatment was concealed using bottles marked A or B. Staff were blind to the treatment allocation. The primary outcome was IBK cumulative incidence over the study period. The secondary outcome was weaning weight. Only the Iowa herd met the criteria for an "at-risk" herd i.e. >15% IBK in unvaccinated calves and M. bovoculi isolation from IBK cases. Analysis was "per-protocol". The cumulative incidence of IBK was 47/105 in vaccinated calves and 49/109 in unvaccinated calves (unadjusted odds ratio=0.99, 95% CI: 0.58-1.70). Weight at weaning did not differ between the vaccinated cohort 148kg (SD: +/-27) and unvaccinated cohort 146kg (SD: +/-26) (unadjusted beta=1.5 and 95% CI: -5.5 to 8.6). Results indicate that the autogenous vaccine was ineffective in this study population.

Spectroscopic and model studies of the Ni-Fe hydrogenase reaction mechanism.

Ni-Fe H2ases exhibit a number of states that have been spectroscopically characterized. The structural information arising from these spectroscopic studies are compared with predictions from theoretical calculations and are used to address aspects of hydrogenase reaction mechanism.

Interaction of Cu,Zn superoxide dismutase with hydrogen sulfide.

Addition of HS- enhanced the O(2-)-scavenging activity of bovine erythrocyte Cu,Zn superoxide dismutase (EC 1.15.1.1) by about twofold. The positive effect was measured using a diverse selection of SOD activity assays, and cannot be an artifact restricted to any single technique. Km values for HS- varied in different assay techniques, but we estimate Km approximately 80 microM HS-. In contrast to HS-, other small molecules tested with SOD either had little effect or were inhibitory. Consumption of HS- and O2- occurred in nearly 1:1 mole ratio. The products were H2O2 and sulfane sulfur, such as either elemental sulfur or polysulfide. Binding of HS- to the enzyme was rapid, with k > 10(7) M-1 s-1. The resulting complex exhibited a Cu-to-S charge-transfer absorbance band at 345 nm and an altered Cu(II) EPR spectrum. Taken together, these observations suggest that HS- binds at the catalytic Cu center of SOD and can be a genuine substrate of the enzyme.

Molecular cloning and characterization of the insecticidal crystal protein gene of Bacillus thuringiensis var. tenebrionis.

The insecticidal crystal protein gene of the coleopteran-toxic Bacillus thuringiensis var. tenebrionis has been isolated, and the nucleotide sequence has been determined. A total DNA library from var. tenebrionis was made in the plasmid vector pUC12. By using a synthetic 27-base oligonucleotide corresponding to a stretch of nine N-terminal amino acids of a tryptic fragment of purified crystal protein of var. tenebrionis as a probe, recombinant colonies were screened by in situ hybridization for the presence of the crystal protein gene. Positive clones obtained from this screening were further tested for toxicity. One recombinant, NSBP544 (which contained a 5.9-kilobase BamHI insert), was toxic to larvae of Colorado potato beetle. Immunoblot analysis revealed that this clone produces two crystal-specific antigens of 65 and 73 kDa as do sporulating var. tenebrionis cells. However, purified crystal inclusions from var. tenebrionis contain a primary peptide component of 65 kDa. A 1932-base-pair open reading frame with a coding capacity of 73,119 Da has been identified by nucleotide sequencing analysis of the cloned crystal protein. In addition, mung bean nuclease mapping indicates that transcription of the crystal protein of var. tenebrionis initiates 130 base pairs upstream from the translational start site. Southern blot analysis using an internal 0.7-kilobase EcoRI fragment of pNSBP544 as a probe revealed that the crystal protein gene is located on a 90-MDa plasmid.

An XAS investigation of product and inhibitor complexes of Ni-containing GlxI from Escherichia coli: mechanistic implications.

Escherichia coli glyoxalase I (GlxI) is a metalloisomerase that is maximally activated by Ni(2+), unlike other known GlxI enzymes which are active with Zn(2+). The metal is coordinated by two aqua ligands, two histidines (5 and 74), and two glutamates (56 and 122). The mechanism of E. coli Ni-GlxI was investigated by analyzing Ni K-edge X-ray absorption spectroscopic (XAS) data obtained from the enzyme and complexes formed with the product, S-D-lactoylglutathione, and various inhibitors. The analysis of X-ray absorption near edge structure (XANES) was used to determine the coordination number and geometry of the Ni site in the various Ni-GlxI complexes. Metric details of the Ni site structure were obtained from the analysis of extended X-ray absorption fine structure (EXAFS). Interaction of S-D-lactoylglutathione (product) or octylglutathione with the enzyme did not change the structure of the Ni site. However, analysis of XAS data obtained from a complex formed with a peptide hydroxamate bound to Ni-GlxI is consistent with this inhibitor binding to the Ni center by displacement of both water molecules. XANES analysis of this complex is best fit with a five-coordinate metal and, given the fact that both histidine ligands are retained, suggests the loss of a glutamate ligand. The loss of a glutamate ligand would preserve the neutral charge on the Ni complex and is consistent with the lack of a significant shift in the Ni K-edge energy in this complex. These data are compared with data obtained from the E. coli Ni-GlxI selenomethionine-substituted enzyme. The replacement of three methionine residues in the native enzyme with selenomethionine does not affect the structure of the Ni site. However, addition of the peptide hydroxamate inhibitor leads to the formation of a complex whose structure as determined by XAS analysis is consistent with inhibitor binding via displacement of both water molecules but retention of both histidine and glutamate ligands. This leads to an anionic complex, which is consistent with an observed 1.7 eV decrease in the Ni K-edge energy. Plausible reaction mechanisms for Ni-GlxI are discussed in light of the structural information available.

The impact of routine CT of the chest on the diagnosis and management of newly diagnosed squamous cell carcinoma of the head and neck.

OBJECTIVE: The purpose of the study was to ascertain how often additional malignant neoplasms are detected on CT scans of the thorax in patients with newly diagnosed squamous cell carcinoma of the head and neck and to determine how often these findings are evident on conventional radiographs of the chest. SUBJECTS AND METHODS: One hundred eighty-nine patients with newly diagnosed squamous cell carcinoma of the head and neck were prospectively examined in a 5-year period. At the time of the initial diagnosis, each patient underwent both chest radiography (posteroanterior and lateral radiographs in 95%, anteroposterior in 5%) and thoracic CT to assess the prevalence of additional primary or metastatic malignant neoplasms of the thorax and upper abdomen. RESULTS: Of the 189 patients studied, 66 showed a total of 73 significant abnormalities on thoracic CT scans of which only 17 abnormalities (23%) were detected on chest radiographs alone. Of these 66 patients, 36 (55%) were found to have one or more primary or metastatic neoplasms. These 36 patients manifested a total of 41 additional primary or secondary malignant neoplasms including 24 cases in which one or more pulmonary nodules were detected, six cases of lymphadenopathy, three hepatic lesions, three bone lesions, two pleural masses, two esophageal masses and one adrenal mass. Of these 41 malignant tumors, 13 (32%) were synchronous primary tumors and 28 were metastases. Only 12 (29%) of the 41 malignant tumors detected by thoracic CT were seen on the chest radiographs. In all patients in which an additional malignant tumor was diagnosed, clinical management was significantly affected, resulting in a modification of the planned surgery or the addition of chemotherapy, radiation therapy, or both. CONCLUSION: A relatively large percentage of patients (19%) with newly diagnosed squamous cell cancer of the head and neck was found to have additional malignant tumors, 32% of which were synchronous primary tumors. The discovery of these additional neoplasms had a major effect on both the therapy and the prognosis of these patients. The combination of a relatively poor detection rate for conventional chest radiography, with only 29% of the malignant tumors detected on CT scans of the chest being seen on chest radiographs, and the high prevalence of disease in this population support the routine inclusion of thoracic CT in these patients.

Treatment of suture line bleeding with a novel synthetic surgical sealant in a canine iliac PTFE graft model.

CoSeal mark surgical sealant (CoSeal) was evaluated for inhibiting suture line bleeding using a canine iliac PTFE graft model. Both iliac arteries of 12 heparinized canines were grafted with PTFE. CoSeal was applied to the suture lines of one graft in each animal. The contra-lateral graft served as a control and bleeding was controlled with gauze and pressure (tamponade). The cross-clamps were removed 30 s following application of CoSeal. Times to hemostasis and volume of blood loss at each graft site were determined. Compared to tamponade control, CoSeal significantly reduced the time to hemostasis (average of 5 min vs. greater than 15 min, p < 0.05) and blood loss (19 g vs. 284 g, p < 0.05). Small amounts of CoSeal were visible grossly or histologically at day 7. Histology showed moderate to marked inflammation in CoSeal sites and moderate inflammation in control sites at day 7. At 30 and 60 days, no CoSeal was visible grossly or histologically. Histology showed moderate inflammation in both CoSeal treated sites and in control sites at day 30 and mild to moderate inflammation in both CoSeal and control sites at day 60. CoSeal significantly reduced the time to hemostasis and blood loss in comparison to tamponade.

Structural examination of the nickel site in chromatium vinosum hydrogenase: redox state oscillations and structural changes accompanying reductive activation and CO binding.

An X-ray absorption spectroscopic study of structural changes occurring at the Ni site of Chromatium vinosum hydrogenase during reductive activation, CO binding, and photolysis is presented. Structural details of the Ni sites for the ready silent intermediate state, SI(r), and the carbon monoxide complex, SI-CO, are presented for the first time in any hydrogenase. Analysis of nickel K-edge energy shifts in redox-related samples reveals that reductive activation is accompanied by an oscillation in the electron density of the Ni site involving formally Ni(III) and Ni(II), where all the EPR-active states (forms A, B, and C) are formally Ni(III), and the EPR-silent states are formally Ni(II). Analysis of XANES shows that the Ni site undergoes changes in the coordination number and geometry that are consistent with five-coordinate Ni sites in forms A, B, and SI(u); distorted four-coordinate sites in SI(r) and R; and a six-coordinate Ni site in form C. EXAFS analysis reveals that the loss of a short Ni-O bond accounts for the change in coordination number from five to four that accompanies formation of SI(r). A shortening of the Ni-Fe distance from 2.85(5) A in form B to 2.60(5) A also occurs at the SI level and is thus associated with the loss of the bridging O-donor ligand in the active site. Multiple-scattering analysis of the EXAFS data for the SI-CO complex reveals the presence of Ni-CO ligation, where the CO is bound in a linear fashion appropriate for a terminal ligand. The putative role of form C in binding H(2) or H(-) was examined by comparing the XAS data from form C with that of its photoproduct, form L. The data rule out the suggestion that the increase in charge density on the NiFe active site that accompanies the photoprocess results in a two-electron reduction of the Ni site [Ni(III) --> Ni(I)] [Happe, R. P., Roseboom, W., and Albracht, S. P. J. (1999) Eur. J. Biochem. 259, 602-608]; only subtle structural differences between the Ni sites were observed.

Use of computed radiography in the study of an historic painting.

The authors demonstrate the use of radiography in the investigation of an historic painting and describe the potential benefits of computed radiography compared with conventional screen-film radiography. The subject for the comparison was a 16 x 19-foot oil-on-canvas painting, Scipio Africanus Freeing Massiva, by Giovanni Battista Tiepolo. Radiographs of the painting were obtained by using a portable, industrial radiographic unit and both conventional screen-film and photostimulable phosphor plate cassettes. For this investigation, computed radiography had a number of advantages over screen-film radiography, largely due to its wider dynamic range and its capabilities for enhancing the digital images with image processing tools such as magnification, edge enhancement, colorization, and airbrushing. The ability to electronically combine images from the large painting into a single composite image file was extremely valuable, as this technique was much less cumbersome and resulted in much higher quality composite images than could be achieved with conventional radiography. An additional advantage of computed radiography includes the capability to easily archive and transmit these images in a digital format for subsequent review.

Isolation, characterization and gene sequence analysis of a membrane-associated 89 kDa Fe(III) reducing cytochrome c from Geobacter sulfurreducens.

Geobacter sulfurreducens is capable of anaerobic respiration with Fe(III) as a terminal electron acceptor via a membrane-bound Fe(III) reductase activity associated with a large molecular mass cytochrome c. This cytochrome was purified by detergent extraction of the membrane fraction, Q-Sepharose ion-exchange chromatography, preparative electrophoresis, and MonoQ ion-exchange chromatography. Spectrophotometric analysis of the purified cytochrome reveals a c-type haem, with no evidence of haem a, haem b or sirohaem. The cytochrome has an M(r) of 89000 as determined by denaturing PAGE, and has an isoelectric point of 5.2 as determined by analytical isoelectric focusing. Dithionite-reduced cytochrome can donate electrons to Fe(III)-nitrilotriacetic acid and synthetic ferrihydrite, thus demonstrating that the cytochrome has redox and thermodynamic properties required for reduction of Fe(III). Analysis using cyclic voltammetry confirmed that the reduced cytochrome can catalytically transfer electrons to ferrihydrite, further demonstrating its ability to be an electron transport mediator in anaerobic Fe(III) respiration. Sequence analysis of a cloned chromosomal DNA fragment revealed a 2307 bp open reading frame (ferA) encoding a 768 amino acid protein corresponding to the 89 kDa cytochrome. The deduced amino acid sequence (FerA) translated from the open reading frame contained 12 putative haem-binding motifs, as well as a hydrophobic N-terminal membrane anchor sequence, a lipid-attachment site and an ATP/GTP-binding site. FerA displayed 20% or less identity with amino acid sequences of other known cytochromes, although it does share some features with characterized polyhaem cytochromes c.

Octopine and nopaline strains of Agrobacterium tumefaciens differ in virulence; molecular characterization of the virF locus.

Octopine and nopaline strains of Agrobacterium tumefaciens were found to differ in virulence on Nicotiana glauca. This difference is due to the absence of a functional virF locus, which is necessary for efficient tumorigenesis on N. glauca, from the nopaline Ti plasmids. Genetic studies and DNA sequence analysis of the virF locus revealed that virF embraces one open reading frame coding for a hydrophilic protein with a molecular mass of 22,437 Da. Transcription of virF is directed from left to right, towards the T region, and is strongly induced by the phenolic compound acetosyringone. We established that virA and virG, two genes known to be essential for induction of the vir regulon, are necessary for acetosyringone-induced virF expression, implying that virF is a member of this vir regulon. Agrobacterium virF mutants can be complemented for tumor induction by co-infection with avirulent Agrobacterium 'helper' strains. We found that such 'helper' strains must express not only the virF gene but also the vir operons virA, virB, virD and virG.

Examination of the nickel site structure and reaction mechanism in Streptomyces seoulensis superoxide dismutase.

Superoxide dismutases are metalloenzymes involved in protecting cells from oxidative damage arising from superoxide radical or reactive oxygen species produced from superoxide. Examples of enzymes containing Cu, Mn, and Fe as the redox-active metal have been characterized. Recently, a SOD containing one Ni atom per subunit was reported. The amino acid sequence of the NiSOD deduced from the nucleotide sequence of the structural gene sodN from Streptomyces seoulensis is reported and has no homology with other SODs. X-ray absorption spectroscopic studies coupled with EPR of the Ni center show that the Ni in the oxidized (as isolated) enzyme is in a five-coordinate site composed of three S-donor ligands, one N-donor, and one other O- or N-donor. This unique coordination environment is modified by the loss of one N- (or O-) donor ligand in the dithionite-reduced enzyme. The NiSOD activity was determined by pulse radiolysis, and a value of kcat = 1.3 x 10(9) M-1 s-1 per Ni was obtained. The rate is pH sensitive and drops off rapidly above pH 8. The results characterize a novel class of metal center active in catalyzing the redox chemistry of superoxide and, when placed in context with other nickel enzymes, suggest that thiolate ligation is a prerequisite for redox-active nickel sites in metalloenzymes.

A tissue sealant based on reactive multifunctional polyethylene glycol.

A rapidly gelling synthetic tissue sealant was developed from tetra-succinimidyl and tetra-thiol-derivatized polyethylene glycol (PEG). The two reagents were dissolved in aqueous buffers at 20% (w/v) solids and sprayed on the tissue site, with the use of a sprayer/mixer device. Good adhesion to collagen membranes, PTFE grafts, and carotid artery was observed in vitro. In a burst test on collagen membranes with a 2-mm orifice defect, the gel sustained fluid pressures of 125 +/- 36 mm Hg (n = 18), fivefold greater than capillary blood pressure and one-half that observed in hypertension. On 0.4-mm-diameter puncture defects in PTFE grafts, pressures of 390-490 mm Hg were sustained, and on 0.6-0.9-mm puncture defects in carotid arteries, pressures of 490 to 840 mm Hg were sustained. In vitro data corresponded to results in vivo, where bleeding in rabbit arteries was stopped immediately in five out of six trials. A significant reduction in time to hemostasis and blood loss, compared to controls, was observed. Carotid artery and subcutaneous implant data in rabbits showed that the formula was compatible with biological tissue. Rapid gelling and effective sealing were dependent on the presence of active succinimidyl ester and thiol groups on PEG. HPLC and chemical substitution methods were useful in predicting whether batches of derivatized PEG would perform satisfactorily.