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50 Pág., In March 2022, following the annual International Committee on Taxonomy of Viruses (ICTV) ratifcation vote on newly proposed taxa, the phylum Negarnaviricota was amended and emended. The phylum was expanded by two new families (bunyaviral Discoviridae and Tulasviridae), 41 new genera, and 98 new species. Three hundred forty-nine species were renamed and/or moved. The accidentally misspelled names of seven species were corrected. This article presents the updated taxonomy of Negarnaviricota as now accepted by the ICTV, This work was supported in part through Laulima Govern ment Solutions, LLC prime contract with the US National Institute of Allergy and Infectious Diseases (NIAID) under Contract No. HHSN272201800013C. J.H.K. performed this work as an employee of Tunnell Government Services (TGS), a subcontractor of Laulima Gov ernment Solutions, LLC under Contract No. HHSN272201800013C. This work was also funded in part by Contract No. HSHQDC 15-C-00064 awarded by DHS S&T for the management and operation of The National Biodefense Analysis and Countermeasures Center, a federally funded research and development center operated by the Battelle National Biodefense Institute (V.W.); and NIH contract HHSN272201000040I/HHSN27200004/D04 and grant R24AI120942 (N.V., R.B.T.). S.S. acknowledges support from the Mississippi Agri cultural and Forestry Experiment Station (MAFES), USDA-ARS pro ject 58-6066-9-033 and the National Institute of Food and Agriculture, US Department of Agriculture, Hatch Project, under Accession Num ber 1021494.

This study aimed to generate data on performance characteristics for 2 real-time TaqMan PCR assays (CSIRO and WOAH WSSV qPCRs) for the purposes of (1) detection of white spot syndrome virus (WSSV) in clinically diseased prawns and (2) detection of WSSV in apparently healthy prawns. Analytical sensitivity of both assays was 2 to 20 genome copies per reaction, and analytical specificity was 100% after testing nucleic acid from 9 heterologous prawn pathogens and 4 prawn species. Results obtained after testing more than 20 000 samples in up to 559 runs with the CSIRO WSSV qPCR and up to 293 runs with the WOAH WSSV qPCR demonstrated satisfactory repeatability for both assays. Both assays demonstrated median diagnostic sensitivity (DSe) 100% (95% CI: 94.9-100%) when testing clinically diseased prawns. When 1591 test results from apparently healthy prawns were analysed by Bayesian latent class analysis, median DSe and diagnostic specificity (DSp) were 82.9% (95% probability interval [PI]: 75.0-90.2%) and 99.7% (95% PI: 98.6-99.99%) for the CSIRO WSSV qPCR and 76.8% (95% PI: 68.9-84.9%) and 99.7% (95% PI: 98.7-99.99%) for the WOAH WSSV qPCR. When both assays were interpreted in parallel, median DSe increased to 98.3 (95% PI: 91.6-99.99%), and median DSp decreased slightly to 99.4% (95% PI: 97.9-99.99%). Routine testing of quantified positive controls by laboratories in the Australian laboratory network demonstrated satisfactory reproducibility of the CSIRO WSSV qPCR assay. Both assays demonstrated comparable performance characteristics, and the results contribute to the validation data required in the WOAH validation pathway for the purposes of detection of WSSV in clinically diseased and apparently healthy prawns.

Optically active defects in 2D materials, such as hexagonal boron nitride (hBN) and transition-metal dichalcogenides (TMDs), are an attractive class of single-photon emitters with high brightness, operation up to room temperature, site-specific engineering of emitter arrays with strain and irradiation techniques, and tunability with external electric fields. In this work, we demonstrate a novel approach to precisely align and embed hBN and TMDs within background-free silicon nitride microring resonators. Through the Purcell effect, high-purity hBN emitters exhibit a cavity-enhanced spectral coupling efficiency of up to 46% at room temperature, exceeding the theoretical limit (up to 40%) for cavity-free waveguide-emitter coupling and demonstrating nearly a 1 order of magnitude improvement over previous work. The devices are fabricated with a CMOS-compatible process and exhibit no degradation of the 2D material optical properties, robustness to thermal annealing, and 100 nm positioning accuracy of quantum emitters within single-mode waveguides, opening a path for scalable quantum photonic chips with on-demand single-photon sources.

In coastal marsh ecosystems, high salinities, anoxic waterlogged soils, and elevated summer temperatures often promote physiological strain that results in only a few tolerant halophytic species. Although not well understood, plant physiological responses to multiple stressors can be complex and may involve intensifying or offsetting reactions. In this study, we investigated physiological responses to combined salinity and high temperature in the coastal marsh graminoid Sporobolus pumilus (syn. Spartina patens). Specifically, we considered changes in plant-water relations and Photosystem II (PSII) behavior (involving chlorophyll [chl] a fluorescence) in heat-shocked plants that were acclimated to different salinities (0, 15, and 30 psu). Higher salinities fostered lower stomatal conductance (g), lower leaf-water potential (Ψleaf) and lower tissue-water content (θ), as well as decreased potential quantum yield (Fv/Fm) and decreased excitation energy capture efficiencies of open reaction centers (Fv’/Fm’). Heat-shocked plants acclimated to freshwater only had decreased Fv/Fm and PSII performance index (PIABS). Interestingly, there were no changes in chl a fluorescent outputs in heat-shocked plants acclimated to moderate salinities, and minimal changes in plants acclimated to high salinities. Approximately 25% of the heat-shocked S. pumilus in freshwater revealed a K-step in their polyphasic chl a fluorescent transients (OJIP procedure); K-steps were not observed in salt-treated plants. This suggests that, for plants residing in freshwater, heat-shock promoted disturbances in the PSII reaction centers and, in some cases, disrupted the oxygen-evolving complex. These PSII disruptions were either absent or less intense in salinity-treated plants, indicating that acclimation to environmental salts may provide PSII thermostability in S. pumilus.

An orthomyxo-like virus was first isolated in 1998 as an incidental discovery from pilchards Sardinops sagax collected from waters off the South Australian coast. In the following 2 decades, orthomyxo-like viruses have been isolated from healthy pilchards in South Australia and Tasmania. In 2006, an orthomyxo-like virus was also isolated from farmed Atlantic salmon Salmo salar in Tasmania during routine surveillance and, again, from 2012 onwards from diseased Atlantic salmon. Using transmission electron microscopy, these viruses were identified as belonging to the family Orthomyxoviridae. To further characterise the viruses, the genomes of 11 viral isolates were sequenced. The open reading frames (ORFs) that encode 10 putative proteins from 8 viral genome segments were assembled from Illumina MiSeq next generation sequencing (NGS) data. The complete genome of a 2014 isolate was also assembled from NGS, RNA-sequencing (RNA-seq) data, that included conserved motifs that shared commonalities with infectious salmon anaemia virus, rainbow trout orthomyxovirus and Influenzavirus A. The presence of 8 viral proteins translated from genome segments was confirmed by mass spectrometric analysis including 2 novel proteins with no known orthologs. Sequence analysis of the ORFs, non-coding regions and proteins indicated that the viruses had minimal diversity and hence were named pilchard orthomyxovirus (POMV), based on the fish host species of its first isolation. The low homology of POMV proteins with previously characterised orthomyxoviruses suggests that POMV is the first virus to be characterised from a new genus within the Orthomyxoviridae. To facilitate more rapid detection and subsequent diagnostic confirmation of POMV infections, TaqMan and conventional nested PCRs were designed.

Samples from multiple animals may be pooled and tested to reduce costs of surveillance for infectious agents in aquatic animal populations. The primary advantage of pooling is increased population-level coverage when prevalence is low (

Expression of fdp, encoding a fasciclin I domain protein important for adherence in the hydrogen-producing bacterium Rhodobacter sphaeroides, was investigated under a range of conditions to gain insights into optimization of adherence for immobilization strategies suitable for H2 production. The fdp promoter was linked to a lacZ reporter and expressed in wild type and in PRRB and PRRA mutant strains of the Prr regulatory pathway. Expression was significantly negatively regulated by Prr under all conditions of aerobiosis tested including anaerobic conditions (required for H2 production), and aerobically regardless of growth phase, growth medium complexity or composition, carbon source, heat and cold shock and dark/light conditions. Negative fdp regulation by Prr was reflected in cellular levels of translated Fdp protein. Since Prr is required directly for nitrogenase expression, we propose optimization of Fdp-based adherence in R. sphaeroides for immobilized biohydrogen production by inactivation of the PrrA binding site(s) upstream of fdp., Highlights • Adherence and immobilization of hydrogen-producing microbes improves H2 yields. • fdp encodes an adherence factor in hydrogen-producer Rhodobacter sphaeroides. • Expression of fdp is negatively regulated by the global Prr regulatory pathway. • Negative regulation by Prr was demonstrated under all growth conditions tested. • Fdp-based adherence may be optimised by inactivating the PrrA binding site(s) of fdp.

Bonamiosis has developed as a problem in Australian native oysters Ostrea angasi since the parasite Bonamia spp. was first detected in Port Phillip Bay, Victoria, in the early 1990s. At that time, large-scale mortalities in both farmed and wild oysters saw the demise of the pilot native oyster culture industry. More recent attempts to farm the species resulted in subclinical infections that progressed over time to clinical disease. The aim of this work was to establish what environmental factors result in the clinical manifestation of disease; determine the diagnostic sensitivity and diagnostic specificity of histopathological examination and a quantitative polymerase chain reaction (qPCR) test for the diagnosis of B. exitiosa infection in clinically diseased farmed native oysters; and calculate the optimal qPCR threshold cycle (CT) epidemiological cut-point for classification of positive and negative cases. After applying a range of stressors to tank-held oysters, results indicated a 58% increased risk (95% CI: 16%, 99%) of a Bonamia-infected oyster dying if the oyster was held at a higher temperature (p = 0.048). Starving and tumbling oysters, in isolation, was not significantly associated with clinical bonamiosis, but a Bonamia-infected oyster was at the greatest risk of death when increased water temperature was combined with both starvation and increased motion (p = 0.02; odds ratio = 3.47). The diagnostic sensitivity and specificity of the World Organisation for Animal Health qPCR protocol were calculated for increasing CT value cut-points from ≤25 to ≤40, with an optimal cut-point identified at ≤34.5 (specificity: 92.2; 95% posterior credible intervals [PCI]: 76.2, 99.8; Sensitivity: 93.5; 95% PCI: 84.7, 99.1).

Wild Acetes sibogae australis from northern Moreton Bay, Australia displaying opacity of the hepatopancreas were sampled and examined histologically, revealing infection by multinucleate plasmodia of a haplosporidian-like parasite in the epithelial cells of the hepatopancreas. A morphological and phylogenetic investigation identified the parasite as a novel species of the order Haplosporida, and the parasite is described as Haplosporidium acetes n. sp. This is the first report of disease caused by a haplosporidian in wild Australian decapod crustaceans, and the first record of haplosporidiosis in sergestid shrimp. Infections of H. acetes were observed in all cell types (R, B, F and E) within the hepatopancreas. Infected epithelial cells became hypertrophied as they filled with haplosporidian parasites and, in heavy infections, caused almost complete displacement of normal hepatopancreas tissue. Although sporulation was not observed, infected jelly prawns appeared terminally diseased. Infections became grossly evident in around 5% of wild prawns during early autumn at a time of year when jelly prawn populations decline rapidly with decreasing water temperatures, however histopathology indicated at least 13% of apparently normal jelly prawns were also infected. Further studies are required in order to determine if this parasite influences jelly prawn population dynamics. In addition, we report co-infection of a novel microsporidian parasite in the Enterocytozoon Group Microsporidia (EGM) infecting nuclei of hepatopancreatic epithelial cells. The microsporidian was phylogenetically distinct from Enterocytozoon hepatopenaei (EHP) known to infect penaeid shrimp in Asia.

Design and reporting quality of diagnostic accuracy studies (DAS) are important metrics for assessing utility of tests used in animal and human health. Following standards for designing DAS will assist in appropriate test selection for specific testing purposes and minimize the risk of reporting biased sensitivity and specificity estimates. To examine the benefits of recommending standards, design information from published DAS literature was assessed for 10 finfish, seven mollusc, nine crustacean and two amphibian diseases listed in the 2017 OIE Manual of Diagnostic Tests for Aquatic Animals. Of the 56 DAS identified, 41 were based on field testing, eight on experimental challenge studies and seven on both. Also, we adapted human and terrestrial-animal standards and guidelines for DAS structure for use in aquatic animal diagnostic research. Through this process, we identified and addressed important metrics for consideration at the design phase: study purpose, targeted disease state, selection of appropriate samples and specimens, laboratory analytical methods, statistical methods and data interpretation. These recommended design standards for DAS are presented as a checklist including risk-of-failure points and actions to mitigate bias at each critical step. Adherence to standards when designing DAS will also facilitate future systematic review and meta-analyses of DAS research literature.

Background The aim of the study was to evaluate the current opinion and clinical decision-making process of international pancreatologists, and to systematically identify key study questions regarding the diagnosis and treatment of chronic pancreatitis (CP) for future research. Methods An online survey, including questions regarding the diagnosis and treatment of CP and several controversial clinical case vignettes, was send by e-mail to members of various international pancreatic associations: IHPBA, APA, EPC, ESGE and DPSG. Results A total of 288 pancreatologists, 56% surgeons and 44% gastroenterologists, from at least 47 countries, participated in the survey. About half (48%) of the specialists used a classification tool for the diagnosis of CP, including the Mayo Clinic (28%), Mannheim (25%), or Buchler (25%) tools. Overall, CT was the preferred imaging modality for evaluation of an enlarged pancreatic head (59%), pseudocyst (55%), calcifications (75%), and peripancreatic fat infiltration (68%). MRI was preferred for assessment of main pancreatic duct (MPD) abnormalities (60%). Total pancreatectomy with auto-islet transplantation was the preferred treatment in patients with parenchymal calcifications without MPD abnormalities and in patients with refractory pain despite maximal medical, endoscopic, and surgical treatment. In patients with an enlarged pancreatic head, 58% preferred initial surgery (PPPD) versus 42% initial endoscopy. In patients with a dilated MPD and intraductal stones 56% preferred initial endoscopic ± ESWL treatment and 29% preferred initial surgical treatment. Conclusion Worldwide, clinical decision-making in CP is largely based on local expertise, beliefs and disbeliefs. Further development of evidence-based guidelines based on well designed (randomized) studies is strongly encouraged.

The accuracy of 3 real-time PCR assays (ORF49, ORF66 and ORF77) and histopathology was evaluated for the purpose of demonstrating or certifying abalone free from Haliotid herpesvirus 1 (AbHV), the causative agent of abalone viral ganglioneuritis. Analytically, all 3 qPCRs showed equivalent limit of detection (20 copies per reaction); however, ORF49 could not detect 2 of the AbHV genotypes. A selection of 1452 archive specimens sourced from apparently healthy abalone populations was screened using all 4 tests. In the absence of a perfect reference standard, a Bayesian latent class analysis was built to estimate diagnostic sensitivity (DSe), diagnostic specificity (DSp) and likelihood ratios of a positive (LR+) and negative test result (LR-) for each individual test and for all possible combinations of test pairs interpreted either in series or in parallel. The pair ORF49/ORF66 interpreted in parallel performed the best both analytically and diagnostically to demonstrate freedom from AbHV in an established population of abalone and to certify individual abalone free from AbHV for trade or movement purposes (DSe = 96.0%, 95% posterior credibility interval [PCI]: 82.6 to 99.9; DSp = 97.7%, 95% PCI: 96.4 to 99.4; LR+ = 41.4, 95% PCI: 27.4 to 148.7; LR- = 0.041, 95% PCI: 0.001 to 0.176). Histopathology showed very poor DSe (DSe = 6.3%, 95% PCI: 2.4 to 13.1) as expected since most infected abalone in the study were likely sub-clinical with limited pathological change. Nevertheless, we recommend histopathology when clinically investigating outbreaks to find potential, new, emerging AbHV genotype(s) that may not be detectable by either ORF49 or ORF66.

Whole dry-cured (country-style) hams from six manufacturers were sliced and the slices randomly allotted into five treatment groups per manufacturer. One treatment group served as a control and slices in the four other treatment groups were inoculated with approximately 105 CFU/g of ham of either Escherichia coli O157:H7, Listeria monocytogenes , a mixture of three Salmonella spp. ( Salmonella typhimurium , Salmonella enteritidis and Salmonella choleraesuis ), or Staphylococcus aureus . All ham slices were vacuum-packaged with half of the packages in each treatment group stored at 25°C and half stored at 2°C. Two packages from each manufacturer for each treatment and storage temperature were examined after storage for 0,7, 14,21, and 28 days. S. aureus was detected in 2 of 60 control slices, Salmonella in 2 of 120, L. monocytogenes in 4 of 120, and E. coli O157:H7 was not detected in any of the 120 control ham slices analyzed before or after storage. Aerobic (26 and 35°C) populations of the control vacuum-packaged hams slices increased (P < 0.05) with storage time and the increase in populations was greater (P < 0.05) in vacuum-packaged hams slices at 25 than at 2°C. The extent of the decreases in populations of the inoculated pathogens during storage of vacuum-packaged dry-cured ham slices varied with manufacturer (P < 0.05) and storage temperature (P < 0.05). Decreases in Salmonella spp. and E. coli O157:H7 populations were greater (P < 0.05) in slices stored at 25 than at 2°C, while decreases in L. monocytogenes were similar at both storage temperatures. Staphylococcus aureus enterotoxin was not detected in either S. aureus -inoculated or control ham slices after storage for 28 days. Survival of these pathogens in vacuum-packaged dry-cured ham slices suggests that contaminated hams may pose a safety risk to consumers if consumed without adequate cooking.

In 2013, a unique seventh yellow head virus genotype (YHV7) was detected in Black Tiger shrimp (Penaeus monodon) broodstock that suffered high mortality following their capture from Joseph Bonaparte Gulf (JBG) in northern Australia. To assist with its diagnosis and assessment of its distribution, prevalence and pathogenicity, YHV7-specific TaqMan real-time qPCR and conventional nested PCR primer sets were designed to ORF1b gene sequences divergent from the other YHV genotypes. Using high (≥108) copies of plasmid (p)DNA controls containing ORF1b gene inserts of representative strains of YHV genotypes 1-7, both PCR tests displayed specificity for YHV7. Amplifications of serial 10-fold dilutions of quantified YHV7 pDNA and synthetic ssRNA showed that both tests could reliably detect 10 genome copies. Pleopods/gills from wild P. monodon sourced from locations in geographically disparate regions across northern Australia as well as 96 juveniles (48 either appearing normal or displaying signs of morbidity) from a commercial pond experiencing mortalities were screened to partially validate the diagnostic capacity of the qPCR test. Based on these data and PCR primer/probe sequence mismatches with other newly identified YHV genotypes, both YHV7-specific PCR tests should prove useful in the sensitive detection and discrimination of this genotype from YHV 2 (gill-associated virus) and YHV6 that can occur in Australian P. monodon, as well as from YHV genotypes currently listed as exotic to Australia.

In an attempt to determine whether or not genetic variants of the Tasmanian strain of Atlantic salmon aquareovirus (TSRV) exist, 14 isolates of TSRV, originating from various locations in Tasmania, covering a 20-year period (1990-2010), obtained from various host species and tissues, and isolated on different cell lines, were selected for this study. Two categories, termed "typical" and "atypical", of variants of TSRV were identified based on preliminary genotypic and phenotypic characterization carried out on these 14 different isolates. In addition, electron microscopic examination indicated the existence of at least three variants based on viral particle size. Finally, this study demonstrated the existence of at least one new variant of TSRV isolates, other than the more commonly isolated typical TSRV isolates, in farmed Tasmanian Atlantic salmon.

In November 2010, a rainbow trout (Oncorhynchus mykiss) hatchery in Victoria reported increased mortality rates in diploid and triploid female fingerlings. Live and moribund fish were submitted for laboratory investigation. All fish showed hyperpigmentation of the cranial half of the body. Histological lesions were seen in all areas of skin examined despite the localised nature of the gross lesions. There was irregular hyperplasia and spongiosis, alternating with areas of thinning and architectural disturbance. Occasionally, particularly in superficial layers of epithe- lium, cells showed large, eosinophilic inclusions that obscured other cellular detail. A small num- ber of fish had necrosis in dermis, subcutis and superficial muscles. Bacteriological culture of skin and gills was negative for all bacterial pathogens, including Flavibacterium columnare, the agent of columnaris disease. Attempts at virus isolation from the skin of affected fish resulted in the development of a cytopathic effect in RTG-2 cell cultures suggestive of the presence of a virus. Negative contrast electron microscopy of cell culture supernatant demonstrated the presence of viral particles with the typical morphology of birnaviruses. Preliminary molecular characterisation identified an aquabirnavirus that differed from both the Tasmanian aquabirnavirus (TABV) and other aquabirnaviruses exotic to Australia. Previous isolates of aquabirnaviruses in Australia and New Zealand have been from healthy fish in a marine environment. This is the first report of an aquabirnavirus isolated from young salmonids at a freshwater hatchery in Australia. The role of the virus in the mortality event on the farm is uncertain as no further deaths attributable to this virus have occurred in the 4 yr since its initial discovery. The virus has been provisionally named Victorian trout aquabirnavirus (VTAB).

One in five adults with intellectual disabilities (ID) known to services display challenging behaviours (CBs), and these individuals are at risk for restrictive practices and poor care. Staff attitudes may contribute to the development and/or maintenance of CBs. We investigated the effectiveness of co-produced Who's Challenging Who? training delivered by people with ID to staff.This study involved a cluster randomised controlled trial (RCT) of Who's Challenging Who? training with follow-up at six and 20 weeks post-randomisation.two staff from each of 118 residential care settings for adults with ID at least one of whom displayed aggressive CB.Self-reported Staff Empathy for people with Challenging Behaviour Questionnaire.intention to treat of all randomised settings. ISCRTN registration: ISRCTN53763600.118 residential settings (including 236 staff) were randomised to either receive training (59 settings) or to receive training after a delay (59 settings). The primary analysis included data from 121 staff in 76 settings (51% of staff, 64% of settings). The adjusted mean difference on the transformed (cubed) Staff Empathy for people with Challenging Behaviour Questionnaire score at the primary end point was 1073.2 (95% CI: -938.1 to 3084.5, P = 0.296) in favour of the intervention group (effect size Cohen's d = .19).This is the first large-scale RCT of a co-produced training course delivered by people with ID. Findings indicated a small positive (but statistically non-significant) effect on increased staff empathy at 20 weeks, and small to moderate effects for staff reported secondary outcomes in favour of the intervention group.

Molecular (PCR) diagnostic tests for the detection and identification of aquareovirus in general, and Tasmanian Atlantic salmon reovirus (TSRV) specifically, were developed, and their diagnostic sensitivity and specificity were determined and compared with virus isolation in cell culture. Intralaboratory and interlaboratory comparison of PCR (conventional hemi-nested RT-PCR & RTqPCR) and virus isolation in cell culture using finfish cell lines, CHSE-214 and EPC, was carried out for the detection and identification of TSRV using field samples of farmed Atlantic salmon Salmo salar , L. from various aquaculture sites around Tasmania. The interlaboratory comparison of diagnostic methods was carried out between two laboratories, AAHL-CSIRO and DPIPWE Tasmania. A total of 144 fish from nine sites (12–33 fish per site) were sampled from two regions of Tasmania (Tamar River estuary in the north and Huon River estuary in the south-east) during late spring to early summer of 2009, and the data were analysed using different statistical approaches. The prevalence of TSRV ranged from 6% to 22% in both regions. All the diagnostic methods (data from both laboratories) had high specificity, while the estimated sensitivity varied between tests with RT-qPCR being the most sensitive (95.2%) method followed by virus isolation and then conventional hemi-nested RT-PCR.

Clonal plant growth is common in aquatic freshwater plants and their success is largely attributed to the sharing of resources (e.g., photosynthates, minerals, and water) within the greater genet. The flow of materials within the clone is largely driven by source-sink dynamics, and in established genets may involve acropetal and/or basipetal flow. During the production of new ramets, however, the mother often provides needed resources to the daughter through acropetal flow. In this study, we consider the role of water sharing in a clonal hydrophyte, Justicia americana, when soil moisture levels around the daughter plant declined. The results of this study indicate that acropetal water sharing is an important function between mother and daughter ramets in this species, as indicated by sustained growth and higher survival in connected daughters residing in water-deprived soils. Interestingly, mother plants, when connected to a drought daughter, began to develop similar xeromorphic features (e.g., greater leaf succulence) even though the parent remained in flooded conditions. We suspect that some physical or chemical signal was conveyed from the daughter to the mother that “forecast” water scarcities, which could prepare the entire genet for potential drought conditions.

The syntheses of CpM(i-PrDAB)2 (M = Nb, Ta; Cp = (C5H5); i-PrDAB = bis-isopropyl-1,4-diazabuta-1,3-diene) are reported. Both show fluxional NMR spectra indicating that the two DAB rings differ. The X-ray crystal structure of CpNb(i-PrDAB)2 shows one ring to be more folded than the other. Density functional calculations have been used to investigate the degree of folding of the chelate ring in the compounds Cp2M(R-DAB), (M = Ti, Zr, Hf, Nb, Ta; R = H, i-Pr; DAB = 1,4-diazabuta-1,3-diene) and CpM(R-DAB)2 (M = Nb and Ta). For Cp2M(R-DAB) the group 4 compounds all have folded rings whereas the Nb and Ta compounds have planar rings. In all compounds the rings are reduced and the folding is driven by the electron number requirements of the metal centre.

In Australia, disease caused by betanodavirus has been reported in an increasing number of cultured finfish since the first report of mortalities in 1990. Partial coat protein gene sequences from the T2 or T4 regions of 8 betanodaviruses from barramundi Lates calcarifer, sleepy cod Oxyeleotris lineolata, striped trumpeter Latris lineata, barramundi cod Cromileptes altivelis, Australian bass Macquaria novemaculata and gold-spotted rockcod Epinephelus coioides from several Australian states were determined. Analysis of the 606 bp nucleotide sequences of the T2 region of 4 isolates demonstrated the close relationship with isolates from the red-spotted grouper nervous necrosis virus (RGNNV) genotype and the Cluster Ia subtype. Comparison of a smaller 289 bp sequence from the T4 region identified 2 distinct groupings of the Australian isolates within the RGNNV genotype. Isolates from barramundi from the Northern Territory, barramundi, sleepy cod, barramundi cod and gold-spotted rockcod from Queensland, and striped trumpeter from Tasmania shared a 96.2 to 99.7% nucleotide identity with each other. These isolates were most similar to the RGNNV genotype Cluster Ia. Isolates from Australian bass from New South Wales and from barramundi from South Australia shared a 98.6% sequence identity with each other. However, these isolates only shared an 85.8 to 87.9% identity with the other Australian isolates and representative RGNNV isolates. The closest nucleotide identity to sequences reported in the literature for the New South Wales and South Australian isolates was to an Australian barramundi isolate (Ba94Aus) from 1994. These 2 Australian isolates formed a new subtype within the RGNNV genotype, which is designated as Cluster Ic.

Objective Bariatric surgery reverses obesity-related comorbidities, including type 2 diabetes mellitus. Several studies have already described differences in anthropometrics and body composition in patients undergoing Roux-en-Y gastric bypass compared with laparoscopic adjustable gastric banding, but the role of adipokines in the outcomes after the different types of surgery is not known. Differences in weight loss and reversal of insulin resistance exist between the 2 groups and correlate with changes in adipokines. Methods Fifteen severely obese women (mean body mass index [BMI]: 46.7 kg/m 2 ) underwent 2 types of laparoscopic weight loss surgery (Roux-en-Y gastric bypass=10, adjustable gastric banding=5). Weight, waist and hip circumference, body composition, plasma metabolic markers, and lipids were measured at set intervals during a 24-month period after surgery. Results At 24 months, patients who underwent Roux-en-Y were overweight (BMI 29.7 kg/m 2 ), whereas patients who underwent gastric banding remained obese (BMI 36.3 kg/m 2 ). Patients who underwent Roux-en-Y lost significantly more fat mass than patients who underwent gastric banding (mean difference 16.8 kg, P P =.003), and levels correlated with weight loss, loss of fat mass, insulin levels, and Homeostasis Model of Assessment 2. Adiponectin correlated with insulin levels and Homeostasis Model of Assessment 2 ( r =−0.653, P =.04 and r =−0.674, P =.032, respectively) in the patients who underwent Roux-en-Y at 24 months. Conclusion After 2 years, weight loss and normalization of metabolic parameters were less pronounced in patients who underwent gastric banding compared with patients who underwent Roux-en-Y gastric bypass. Our findings require confirmation in a prospective randomized trial.

A laparoscopically implantable electrical device that intermittently blocks both vagi near the esophagogastric junction led to significant excess weight loss (EWL) in an initial clinical trial in obese patients. The study objective was to optimize therapy algorithms and determine the EWL achieved with a second-generation device at university hospitals in Australia, Norway, and Switzerland.Data acquired during the initial clinical trial were analyzed and subsequently used to select alternative electrical algorithms. In the second trial, vagal blocking using one selected therapy algorithm was initiated 2 weeks after implanting the second-generation device. The patients were followed up for 6 months to assess the EWL and safety, including adverse events.In the initial clinical trial, vagal blocking algorithm durations of 90-150 s were associated with greater EWL compared with either shorter or longer algorithm durations (P.01). The second trial enrolled 27 patients (mean body mass index 39.3+/-.8 kg/m2) to evaluate a 120-s blocking algorithm. At 6 months, greater EWL was achieved (22.7%+/-3.1%, n=24) compared with the initial study and first-generation device (14.2%+/-2.2%, n=29, P=.03). In both trials, an association was found between the number of 90-150-s algorithms delivered daily and greater EWL (P=.03). No deaths, unanticipated device-related adverse events, or medically serious adverse events were associated with the device.This second-generation vagal blocking device, using a therapy algorithm of 120-s duration, resulted in a clinically acceptable safety profile and significantly greater EWL compared with the first-generation device delivering a wider range of therapy algorithm durations.

Five years (2000–04) of spatially complete snow-free land surface albedo data have been produced using high-quality-flagged diffuse bihemispherical (white sky) and direct-beam directional hemispherical (black sky) land surface albedo data derived from observations taken by the Moderate-Resolution Imaging Spectroradiometer (MODIS) instrument aboard the NASA Terra satellite platform (MOD43B3, collection 4). In addition, a spatially complete snow-free aggregate albedo climatological product was generated. These spatially complete products were prepared using an ecosystem-dependent temporal interpolation technique that retrieves missing data within 3%–8% error. These datasets have already been integrated into research and operational projects that require snow-free land surface albedo. As such, this paper provides details regarding the spatial and temporal distribution of the filled versus the original MOD43B3 data. The paper also explores the intra- and interannual variation in the 5-yr data record and provides a qualitative comparison of zonal averages and annual cycles of the filled versus the original MOD43B3 data. The analyses emphasize the data’s inter- and intraannual variation and show that the filled data exhibit large- and small-scale phenological behavior that is qualitatively similar to that of the original MOD43B3. These analyses thereby serve to showcase the inherent spectral, spatial, and temporal variability in the MOD43B3 data as well as the ability of the fill technique to preserve these unique regional and pixel-level phenological characteristics.

The study objective was to define muscle metabolic and cardiovascular changes after surgical intervention in clinically severe obese patients.Obesity is a state of metabolic dysregulation that can lead to maladaptive changes in heart and skeletal muscle, including insulin resistance and heart failure. In a prospective longitudinal study, 43 consecutive patients underwent metabolic profiling, skeletal muscle biopsies, and resting echocardiograms at baseline and 3 and 9 months after bariatric surgery.Body mass index decreased (mean changes, 95% confidence interval [CI]): 7.7 kg/m(2) (95% CI, 6.70-8.89) at 3 months and 5.6 kg/m(2) (95% CI, 4.45-6.80; P.0001) at 9 months after surgery, with restoration of insulin sensitivity and decreases in plasma leptin at the same time points. Concurrent with these changes were dramatic decreases in skeletal muscle transcript levels of stearoyl coenzyme-A desaturase and pyruvate dehydrogenase kinase-4 at 3 and 9 months (P.0001, for both) and a significant decrease in peroxisome proliferation activated receptor-alpha-regulated genes at 9 months. Left ventricular relaxation impairment, assessed by tissue Doppler imaging, normalized 9 months after surgery.Weight loss results in the reversal of systemic and muscle metabolic derangements and is accompanied by a normalization of left ventricular diastolic function.

This study describes the electromyographic characteristics of the gastrointestinal tract of prairie dogs, and examines the effects of a 1.2% cholesterol diet on intestinal myoelectrical activity. Twelve prairie dogs were implanted with eight bipolar electrodes in the stomach and small bowel. Recordings were obtained during fasting (n = 12), gastric instillation of food (n = 7), and chronic cholesterol feeding for 3 weeks (n = 7). Gastric slow waves had a frequency per minute of 4.9 ± 1. Intestinal slow waves had a frequency per minute of 19.1 ± 2 in the duodenum, and of 16.8 ± 1 in the ileum. The migrating myoelectric complex (MMC) was identified, with an interval between two consecutive phases I of 145 ± 33 minutes. The delivery of diets of equal weight into the stomach interrupted fasting activity for approximately 3 hours. This was followed by a duodenal phase III and return of the fasted pattern. The interval between the first two postprandial phases I of the MMC not only was shorter than in the fasted state, but also was shorter after cholesterol diet than after regular diet (89 ± 33 min versus 116 ± 24 min, p ≤ 0.05). Chronic feeding of cholesterol diet did not affect the length of the MMC. In conclusion, the gastrointestinal tract of prairie dogs exhibits slow waves, spike bursts, and an MMC that is interrupted by feeding. High-cholesterol diet does not induce significant changes in the pattern of electrical activity.

Experiments were designed to study the effects of acute pancreatitis, induced by bile duct ligation, on the contractility of longitudinal muscle from the jejunum of rats. Forty animals were divided randomly into 5 groups, 8 in each. In three groups, the common bile duct was ligated at its entrance into the duodenum (low ligation), and their jejunal longitudinal muscle contractility was studied at 24, 48 or 72 hours after operation. In the fourth group, the common bile duct was ligated above the pancreas (high ligation). These animals were studied 72 hours after operation. In the fifth group, sham operation was performed as a control. At the time of study, two strips of longitudinal muscle were peeled from a segment of the jejunum from each animal, and were mounted in organ baths. Muscle contractions in response to carbachol (10-7-10-4m) and to KCL (30 mM) were measured, correlated to the cross-sectional area of the muscle strips, and expressed as stress. Animals with low ligation developed histological evidence of acute pancreatitis. Maximal stress to carbachol of intestinal muscle from these animals decreased progressively with time after operation. Forty-eight and 72 hours following low ligation, Maximal stresses in response to carbachol were 488 ± 33 g cm-2 and 438 ± 28 g cm-2, values significantly lower than those after sham operation (611 ± 24 g cm-2). In contrast, median effective concentrations of carbachol (D-50s) were not significantly different among groups. The response to KCl, although lower than that to carbachol in each group, did not differ among groups. Animals with high ligation, although showing signs of bile stasis, did not develop histological evidence of pancreatitis, and maximal stresses to either carbachol or KCl developed by muscle from these animals did not differ significantly from control. We conclude that low ligation with the induction of pancreatitis leads to a decreased contractility of the jejunum. Also, the impairment responsible for the decreased contractility may reside in an excitation-contraction pathway step initiated by carbachol but not by KCl.

A new medical device uses high-frequency electrical algorithms to create intermittent vagal blocking (VBLOC therapy). The aim is to assess the effects of vagal blocking on excess weight loss (EWL), safety, dietary intake, and vagal function.An open-label, 3-center study was conducted in obese subjects (body mass index [BMI] 35-50 kg/m(2)). Electrodes were implanted laparoscopically on both vagi near the esophagogastric junction to provide electrical block. Patients were followed for 6 months for body weight, safety, electrocardiogram, dietary intake, satiation, satiety, and plasma pancreatic polypeptide (PP) response to sham feeding. To specifically assess device effects alone, no diet or exercise programs were instituted.Thirty-one patients (mean BMI, 41.2 +/- 1.4 kg/m(2)) received the device. Mean EWL at 4 and 12 weeks and 6 months after implant was 7.5%, 11.6%, and 14.2%, respectively (all P.001); 25% of patients lost25% EWL at 6 months (maximum, 36.8%). There were no deaths or device-related serious adverse events (AEs). Calorie intake decreased by30% at 4 and 12 weeks and 6 months (all Por= .01), with earlier satiation (P.001) and reduced hunger (P = .005). After 12 weeks, plasma PP responses were suppressed (20 +/- 7 vs 42 +/- 19 pg/mL). Average percent EWL in patients with PP response25 pg/mL was double that with PP response25 pg/mL (P = .02). Three patients had serious AEs that required brief hospitalization, 1 each for lower respiratory tract, subcutaneous implant site seroma, and Clostridium difficile diarrhea.Intermittent, intra-abdominal vagal blocking is associated with significant EWL and a desirable safety profile.

Complete and transparent reporting of key elements of diagnostic accuracy studies for infectious diseases in cultured and wild aquatic animals benefits end-users of these tests, enabling the rational design of surveillance programs, the assessment of test results from clinical cases and comparisons of diagnostic test performance. Based on deficiencies in the Standards for Reporting of Diagnostic Accuracy (STARD) guidelines identified in a prior finfish study (Gardner et al. 2014), we adapted the Standards for Reporting of Animal Diagnostic Accuracy Studies-paratuberculosis (STRADAS-paraTB) checklist of 25 reporting items to increase their relevance to finfish, amphibians, molluscs, and crustaceans and provided examples and explanations for each item. The checklist, known as STRADAS-aquatic, was developed and refined by an expert group of 14 transdisciplinary scientists with experience in test evaluation studies using field and experimental samples, in operation of reference laboratories for aquatic animal pathogens, and in development of international aquatic animal health policy. The main changes to the STRADAS-paraTB checklist were to nomenclature related to the species, the addition of guidelines for experimental challenge studies, and the designation of some items as relevant only to experimental studies and ante-mortem tests. We believe that adoption of these guidelines will improve reporting of primary studies of test accuracy for aquatic animal diseases and facilitate assessment of their fitness-for-purpose. Given the importance of diagnostic tests to underpin the Sanitary and Phytosanitary agreement of the World Trade Organization, the principles outlined in this paper should be applied to other World Organisation for Animal Health (OIE)-relevant species.

Previous research has shown that beef quality decreased with the age of cattle. In this study, beef trimmings from nine mature cows ( n = 9), equally representing three animal age groups (2–4, 6–8, and 10–12 yr), were restructured into steaks formulated with propyl gallate, alone or in combination with a beefy flavoring agent, to enhance palatability and stability during 6 months of frozen storage at −29 °C. Lipid oxidation, rancidity, and loss of beefy flavor in restructured steaks during extended storage were reduced by propyl gallate. The beefy flavoring agent inclusion masked mature, forage-fed beef off-flavors, intensified beefy flavor, and improved steak tenderness, juiciness and cooking yield. Thus, the combination of propyl gallate and beefy flavoring offers an effective means to enhance the palatability and storage stability of restructured beef prepared from mature cows.

In this paper, we present five-year (2000–2004) climatological statistics of Northern Hemisphere spectral white-sky albedo for the 16 International Geosphere–Biosphere Program (IGBP) ecosystem classes when accompanied by the presence of snow on the ground. These statistics are obtained using validated, high quality Moderate Resolution Imaging Spectroradiometer (MODIS) land surface albedo (MOD43B3) data flagged as snow in the associated Quality Assurance (QA) fields. Near Real-Time Ice and Snow Extent (NISE) data are used as an additional discriminator of snow extent. Statistics are provided for the first seven MODIS bands, ranging from 0.47 to 2.1 μm, and for three broadbands, 0.3–0.7, 0.3–5.0 and 0.7–5.0 μm. The statistics demonstrate that each ecosystem classification has a discernible spectral albedo signature when accompanied by snow on the ground. This indicates that winter canopy and the underlying surface radiative properties are impacted by the presence of snow overlying these surfaces. For example, the 0.47 μm albedo of winter snow-free evergreen needleleaf forests increases from 0.03 to 0.36 in the presence of snow, compared to an increase of 0.04 to 0.76 for croplands. In general, the albedo of snow-covered ecosystems with some winter canopy has lower albedos than ecosystems with little to no winter canopy; for example the 0.47 μm albedo of snow-covered mixed forests is 0.39 compared to 0.87 for barren/deserts and 0.95 for permanent snow. These statistics can be used within land surface models in a stand-alone mode, to prescribe albedo values in atmospheric General Circulation Models (GCMs), or be incorporated into research and operational projects. They are intended to provide researchers with representative spectral albedo values of IGBP ecosystems in the presence of snow that are derived from validated satellite data.

Several Australian betanodaviruses (NNVs) and other, exotic NNVs were used in a study of the performance of a nested PCR test in common use in Australia and New Zealand. This test for the detection of NNVs, designated the ANZSDP VNN Nested PCR, was compared with a currently available commercial PCR kit for NNV detection. With respect to the endemic viruses, the ANZSDP was pan-specific for the NNVs tested, while the commercial PCR kit failed to detect the South Australian strain of barramundi Lates calcarifer NNV. With respect to the relative sensitivities of the two tests, in general, the ANZSDP appeared more sensitive than the commercial test. The exception was the Australian bass Macquaria novemaculeata NNV isolate, which was detected at lower levels by the commercial test. Among the exotic NNVs, both tests detected red-spotted grouper nervous necrosis virus and barfin flounder nervous necrosis virus, but neither test detected striped jack nervous necrosis virus (SJNNV). Further investigation revealed mismatches between SJNNV sequences and the nested primers used in the ANZSDP.

Viruses of the genus Megalocytivirus have not been detected in wild populations of fish in Australia but circulate in imported ornamental fish. In 2012, detection of a megalocytivirus in healthy platys Xiphophorus maculatus was reported from a farm in Australia during surveillance testing as part of a research project undertaken at the University of Sydney. Confirmatory testing of the original samples at the AAHL Fish Diseases Laboratory verified the presence of an infectious spleen and kidney necrosis virus (ISKNV)-like virus. Additional sampling at the positive farm confirmed the persistence of the virus in the platys, with 39 of 265 (14.7%) samples testing positive. Comparison of 3 separate gene regions of the virus with those of ISKNV confirmed the detection of a virus indistinguishable from ISKNV. Subsequently, ISKNV was also detected in a range of imported ornamental fish from several countries between 2013 and 2014, by screening with real-time PCR and confirmation by conventional PCR and sequence analysis. Accordingly, the current importation of live ornamental fish acts as a potential perpetual source for the establishment of ISKNV viruses within Australia. The testing of the farmed and imported ornamental fish verified the utility of the probe-based real-time PCR assay for screening of ornamental fish for Megalocytivirus.

Tasmanian aquabirnaviruses (TABVs) have been isolated intermittently since 1998 from healthy Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss farmed in Macquarie Harbour, Tasmania, Australia. However, beginning in 2011, TABVs have been isolated from rainbow trout in association with mortality events. To determine if recent molecular changes in TABV were contributing to increased mortalities, next generation sequencing was undertaken on 14 TABVs isolated from 1998 to 2013. Sequencing of both genome segments and analysis of the 5 viral proteins they encode revealed that minimal changes had occurred in the past 15 yr. Of the amino acid changes detected only 1, alanine to aspartic acid at position 139 of the minor structural VP3 protein, was unique to the recent disease events. The most dramatic changes observed were in the length of the non-structural VP5 protein varying from 43 to 133 amino acids. However, the amino acid substitution in VP3 and variable VP5 length were unlikely to have resulted in increased TABV pathogenicity. The genome of a novel Australian aquabirnavirus, Victorian trout aquabirnavirus (VTAB) was also sequenced and compared to TABV isolates.

Recent production of land surface anisotropy, diffuse bihemispherical (white-sky) albedo, and direct-beam directional hemispherical (black-sky) albedo from observations acquired by the Moderate Resolution Imaging Spectroradiometer (MODIS) instruments aboard the National Aeronautics and Space Administration's Terra and Aqua satellite platforms have provided researchers with unprecedented spatial, spectral, and temporal information on the land surface's radiative characteristics. Cloud cover, which curtails retrievals, and the presence of ephemeral and seasonal snow limit the snow-free data to approximately half the global land surfaces on an annual equal-angle basis. This precludes the MOD43B3 albedo products from being used in some remote sensing and ground-based applications, climate models, and global change research projects. An ecosystem-dependent temporal interpolation technique is described that has been developed to fill missing or seasonally snow-covered data in the official MOD43B3 albedo product. The method imposes pixel-level and local regional ecosystem-dependent phenological behavior onto retrieved pixel temporal data in such a way as to maintain pixel-level spatial and spectral detail and integrity. The phenological curves are derived from statistics based on the MODIS MOD12Q1 IGBP land cover classification product geolocated with the MOD43B3 data. The resulting snow-free value-added products provide the scientific community with spatially and temporally complete global white- and black-sky surface albedo maps and statistics. These products are stored on 1-min and coarser resolution equal-angle grids and are computed for the first seven MODIS wavelengths, ranging from 0.47-2.1 /spl mu/m and for three broadband wavelengths 0.3-0.7, 0.3-5.0, and 0.7-5.0 /spl mu/m.

Multiple pressures have stimulated the development of curricula to teach fundamental technical skills to surgeons in a laboratory setting. These include reduced resident work hours, increasing costs of operating room time, and the public and payers’ focus on medical errors and the ethics of learning basic skills on patients. In response to these demands, laparoscopic simulators have been developed using inanimate box trainers or computer-based virtual reality platforms.1,2 The goals of these simulator-based curricula are to provide an opportunity to learn and practice basic skills in a relaxed and inexpensive environment to attain a basic level of technical facility that can be transferred from the laboratory to the operating room environment. Laparoscopy has been an area where simulator curricula have attracted much interest because unique skills had to be learned not only by surgeons in training but also by surgeons in practice. This latter group had to develop a strategy to acquire novel skills and incorporate these skills into their clinical practice. Since simulator training requires an investment in both the equipment and time required for training, it is important that this investment be justified by providing proof of the value of simulators. The process to develop and prove the value of the MISTELS (McGill Inanimate System for Training and Evaluation of Laparoscopic Skills) physical laparoscopy simulator has followed a stepwise progression. First, the skills unique to laparoscopic surgery were identified, modeled into exercises that could be carried out in a physical simulator, and a measuring system (metrics) was developed for each exercise, providing a quantitative and objective assessment of performance based on efficiency and precision. Next, the metrics were evaluated for reliability and validity. The most important aspect of validity assessment was to evaluate the relationship of technical skill measured in the simulator to skill in the operating room. If this relationship was found to be robust, simulator performance could then be used to predict performance in the operating room. Once these steps had been completed, the simulator system could then be assessed as a means to verify that laparoscopic technical skill had reached a level thought necessary for the safe performance of basic laparoscopic surgery. In other words, we sought to determine whether the MISTELS score could be used as a summative assessment tool to separate a group that was considered competent from one that would not be considered competent purely from the technical-skill point of view. If so, the sensitivity, specificity, positive and negative predictive values could be determined. The purpose of this paper is to review the process used to develop the MISTELS physical laparoscopy simulator and to summarize the data accumulated over a series of experiments to prove its value as an effective tool to teach and evaluate the fundamental skills required in laparoscopic surgery. These new data are put in context with previously published preliminary data on the MISTELS system.

Beef trimmings from cattle finished on forage or grain were restructured into steaks to enhance palatability. Steaks were treated with propyl gallate with or without a beefy flavoring agent, stored at −29 °C, and analyzed after 0, 1, 3, and 6 months. The strong grassy flavor of forage-finished beef steaks, detected by a sensory panel, was masked by the beefy flavoring agent, rendering the beef more acceptable by consumers. Propyl gallate retarded lipid oxidation and rancidity development in steaks during extended frozen storage. Microbial populations decreased while color scores, cooking yield, and binding strength of steaks exhibited only minor changes during storage. Thus, the combination of antioxidant and flavoring agents with the muscle restructuring technology provides an effective means to enhance the palatability and storage stability of beef from forage-fed cattle.

Aerosol processes that produce titania particles by reacting gaseous precursors (such as titanium tetrachloride) initially must mix the precursor into the oxidizer at elevated temperatures to initiate the formation of product. Oftentimes the rate of reaction is sufficiently large as to be mixing limited. Thus the rate of mixing of the reacting species will control the chemistry and morphological properties of the particles that are produced. The interplay between mixing, nucleation, and growth in these systems is difficult to observe experimentally due to the small time scales that are involved and the spatial limitations of most diagnostics. An alternative approach is direct numerical simulation (DNS). DNS refers to a class of numerical solutions of the three-dimensional time-dependent governing equations for a particular system in which no turbulence modeling assumptions are made. To within the precision of the numerical algorithm, DNS can be thought of as a numerical experiment. Here we apply DNS to th...

Aim To examine the expression of oestrogen receptors α and β (ERα and ERβ) and their regulation by 17β-oestradiol (E2) in stromal cells and adipocytes from human subcutaneous (s.c.) and omental (o.m.) adipose tissue. Methods Subcutaneous and o.m. abdominal adipose tissues were obtained from 10 women (mean age 63.5 ± 4.8 years; mean weight 75.6 ± 6.7 kg) undergoing elective or cosmetic surgery. Immunohistochemistry and RT-PCR analysis were used to detect the presence of ERα and ERβ. The regulation of ERα and ERβ by E2 (10−7 M to 10−9 M) was examined using Western immunoblotting analysis in both s.c. and o.m. stromal cells and mature adipocytes cultured in serum-free, phenol red-free medium. Results Immunostaining of s.c. and o.m. adipose tissue showed that the ER subtypes were localized predominantly within the nucleus. Western analysis demonstrated that E2 treatments differentially altered ERα and ERβ expression in s.c. and o.m. adipocytes. In s.c. and o.m. stromal cells, E2 (10−8 M) produced a significant up regulation relative to control of 66 kDa ERα (s.c.:1.87 ± 0.22; o.m.:1.97 ± 0.17; p

Common bile duct stones are generally classified as primary or secondary stones based on the locations of origin. The vast majority of the stones found in the biliary tree are secondary stones. The current review discusses the pathogenesis and presentations of primary and secondary biliary stones. Based on discussion of disease pathogenesis and presentation, recommendations for the evaluation and management of common and uncommon disease processes associated with choledocholithiasis are proposed.

Background: The cause of postinjury intestinal mucosal barrier disruption remains obscure. The present study examines the hypothesis that the bacterial toxin formyl-methionyl leucyl phenylalanine (FMLP) plays an initial role in this process. Methods: Mucosal permeability to fluoroscein isothiocyanate-labeled dextran (4,400 molecular weight) was measured in perfused distal rat ileum with and without FMLP. Dextran and myeloperoxidase appearance in the lumenal perfusate was assessed in response to surrogates of traumatic stress: ischemia/reperfusion, total abdominal irradiation, and total parenteral nutrition. Recovery of FMLP in the effluent of static closed and perfused ileal loops was determined by mass spectrometry. Release of mast cell mediators in the presence of FMLP was determined in ileal everted sacs. Results: Seventy-five percent of FMLP was recovered in perfusion effluent in contrast to 5% in closed loops. There was a transient increase in ileal permeability in FMLP/perfused, untreated rats, and in ischemia/reperfusion and total parenteral nutrition treated rats that was recorded with a concomitant increment in myeloperoxidase (inflammatory marker) in all experimental models except irradiated rats, which were unresponsive to FMLP. FMLP responsiveness was associated with a significant rise in release of serotonin (mast cell mediator). Conclusion: These results suggest that mast cells and other resident inflammatory cells within the gut wall are involved in FMLP-induced changes in mucosal barrier permeability and raise the possibility that under conditions of traumatic stress, proinflammatory mediators within the gut wall might be activated by toxic factors in the gut lumen.

Temporal changes in tumor necrosis factor-α (TNF-α) and nitric oxide synthase 2 (NOS 2) were evaluated in segments of duodenum, jejunum, and ileum removed from male Sprague-Dawley rats 30, 60, 120, 180, and 240 min after lipopolysaccharide (LPS), 5 mg/kg, intreperitoneally. Plasma was assayed for TNF-α and for nitrate/nitrite (NOx). Intestinal and plasma TNF-α were elevated by 60 min after LPS and were back to control levels by 180 min. For control rats, NOS 2 was detected in the ileum, but not in the duodenum or the jejunum. In rats treated with LPS, NOS 2 was detected in all areas of the intestine at 120 min and was greatest at 240 min. Plasma NOx was elevated at 120 min and continued to increase to 240 min. The time course of changes in intestinal TNF-α and NOS 2 were similar to those reported for other tissues and suggest that the early and late actions of the LPS on the intestine may involve both mediators.

Physical changes in chicken gastrocnemius myofibrils incubated in 0.1 to 1.0 M NaCl solutions with or without 10 mM ortho-(P), pyro-(PP), tripoly-(TPP) or hexameta- (HMP) phosphate at pH 6.0 were examined by phase-contrast microscopy, electrophoresis, and solubility. PP and TPP performed similarly in promoting protein extraction, P had no apparent effect, and HMP exhibited an intermediate effect. PP, TPP, and HMP treatments markedly improved protein solubility in 0.3 and 0.4 M NaCl through the release of myosin, but the phosphate effect diminished in ≥ 0.6 M NaCl. Overall, phosphates influenced the ultrastructure of myofibrils and extraction of their constituents in the order: PP ∼ TPP > HMP > P ∼ nonphosphate control.