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Significance: NSCLC is the leading cause of cancer death due, in part, to a lack of active therapies in advanced disease. We demonstrate that combination therapy with a proteasome inhibitor, BH3-mimetic, and chemotherapy is an active precision therapy in NSCLC cells and tumors expressing Onc-p53 alleles., (©2024 The Authors; Published by the American Association for Cancer Research.)

Objectives: In this study, we investigated whether cardioprotective properties of sevoflurane were expressed in patients with rheumatic heart disease undergoing heart valve surgery under cardiopulmonary bypass (CPB)., Methods: Fifty patients with rheumatic heart disease undergoing heart valve surgery under CPB were randomly assigned to receive total anesthesia with sevoflurane or propofol during surgery. Except for this, anesthetic and surgical management was the same in all patients. The primary outcomes were postoperative high-sensitive cardiac troponin T (hs-cTnT) and creatine kinase-myocardial band (CK-MB) release. The secondary outcomes were hemodynamic events and short-term clinical outcomes (within 30 days after surgery)., Results: The plasma concentrations of hs-cTnT at 24-hour and CK-MB from 6-hour to 48-hour in the sevoflurane group were lower than those in the control group (the propofol group). After aortic unclamping, heartbeat recovery was faster and the rate of sinus rhythm was higher in the sevoflurane group than in the control group. Moreover, a lower proportion of pacemaker use and the need for intraoperative and postoperative inotropes were also found in the sevoflurane group. Nevertheless, there were no differences between the two groups regarding short-term clinical outcomes (durations of mechanical ventilation, intensive care unit stay, hospital stay, morbidity, and mortality rates)., Conclusion: Sevoflurane administered during the entire anesthetic procedure had a myocardial protective effect with less evidence of myocardial damage in the first 48-hour postoperatively but short-term clinical outcomes were not significantly different when compared with the control group in patients with rheumatic heart disease undergoing heart valve surgery under CPB., Competing Interests: Conflicts of interest: All authors have no conflict of interest to declare., (© 2024 Saudi Heart Association.)

Dendrites are injured in a variety of clinical conditions such as traumatic brain and spinal cord injuries and stroke. How neurons detect injury directly to their dendrites to initiate a pro-regenerative response has not yet been thoroughly investigated. Calcium plays a critical role in the early stages of axonal injury detection and is also indispensable for regeneration of the severed axon. Here, we report cell and neurite type-specific differences in laser injury-induced elevations of intracellular calcium levels. Using a human KCNJ2 transgene, we demonstrate that hyperpolarizing neurons only at the time of injury dampens dendrite regeneration, suggesting that inhibition of injury-induced membrane depolarization (and thus early calcium influx) plays a role in detecting and responding to dendrite injury. In exploring potential downstream calcium-regulated effectors, we identify L-type voltage-gated calcium channels, inositol triphosphate signaling, and protein kinase D activity as drivers of dendrite regeneration. In conclusion, we demonstrate that dendrite injury-induced calcium elevations play a key role in the regenerative response of dendrites and begin to delineate the molecular mechanisms governing dendrite repair., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Insulin is secreted in pulses from pancreatic beta-cells, and these oscillations maintain fasting plasma glucose levels within a narrow normal range. Within islets, beta-cells exhibit tight synchronization of regular oscillations. This control circuit is disrupted in type 2 diabetes, and irregularities in pulse frequency and amplitude occur. The prevalence of type 2 diabetes is three times higher in American Indian and Native Alaskans compared to Whites, and their genetic ancestry is associated with low beta-cell function. Obesity in this population compounds their vulnerability to adverse outcomes. The purpose of this article is to review insulin secretion and action and its interaction with race. We also present the results from a 6-month retrospective chart review of metabolic outcomes following intravenous physiologic hormone administration to 10 Native Americans. We found reductions in hemoglobin A1C (baseline: 9.03% ± 2.08%, 6 months: 7.03% ± 0.73%, p = 0.008), fasting glucose (baseline: 176.0 ± 42.85 mg/dL, 6 months: 137.11 ± 17.05 mg/dL, p = 0.02), homeostatic model assessment of insulin resistance (baseline: 10.39 ± 4.66, 6 months: 7.74 ± 4.22, p = 0.008), and triglycerides (baseline: 212.20 ± 101.44, 6 months: 165.50 ± 76.48 mg/dL, p = 0.02). Physiologic hormone administration may improve components of the metabolic syndrome. The therapy warrants investigation in randomized controlled trials., (© 2023 World Obesity Federation.)

Silicon carbide has a planar two-dimensional structure; therefore it is a potential material for constructing twisted bilayer systems for applications. In this study, DFT calculations were performed on four models with different twist angles. We chose angles of 21.8°, 17.9°, 13.2°, and 5.1° to estimate the dependence of the electronic and phononic properties on the twist angle. The results show that the band gap of bilayer SiC can be changed proportionally by changing the twist angle. However, there are only small variations in the band gaps, with an increment of 0.24 eV by changing the twist angle from 5.1° to 21.8°. At four considered twist angles, the band gaps decrease significantly when fixing the structure of each layer and pressing the separation distance down to 3.5 Å, 3.0 Å, 2.7 Å, and 2.5 Å. A noteworthy point is that the pressing also makes the band linearly smaller at a certain rate regardless of the twist angles. Meanwhile, the phonon bands are not affected by the value of the twist angle. The optical bands are between 900 cm -1 and 1100 cm -1 and the acoustic bands are between 0 cm -1 and 650 cm -1 at four twist angles., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Ferroptosis is a regulated cell death modality that occurs upon iron-dependent lipid peroxidation. Recent research has identified many regulators that induce or inhibit ferroptosis; yet, many regulatory processes and networks remain to be elucidated. In this study, we performed a chemical genetics screen using small molecules with known mode of action and identified two agonists of the nuclear receptor Farnesoid X Receptor (FXR) that suppress ferroptosis, but not apoptosis or necroptosis. We demonstrate that in liver cells with high FXR levels, knockout or inhibition of FXR sensitized cells to ferroptotic cell death, whereas activation of FXR by bile acids inhibited ferroptosis. Furthermore, FXR inhibited ferroptosis in ex vivo mouse hepatocytes and human hepatocytes differentiated from induced pluripotent stem cells. Activation of FXR significantly reduced lipid peroxidation by upregulating the ferroptosis gatekeepers GPX4, FSP1, PPARα, SCD1, and ACSL3. Together, we report that FXR coordinates the expression of ferroptosis-inhibitory regulators to reduce lipid peroxidation, thereby acting as a guardian of ferroptosis., (© 2023. The Author(s).)

The escalation of regulatory obligations in order to reshape and implement eco-friendly operations make it essential for firms to execute the related strategies and enhance their position in a competitive market. Scholars and practitioners, thereby, have aligned their search in the area of sustainability to get to know regarding activities that might improve the outcomes of business practices. To contribute in a particular area, the study attempts to explore the critical role of knowledge management, technological advancement, and green supply chain (SC) management on SBP in the presence of a moderator-leadership. China is used as a study sample, and the study model was evaluated through structural equation modelling. Statistical findings indicate that knowledge management, technological advancement, and green SC management have a positive influence on SBP in China. The findings reveal that leadership support significantly moderates knowledge management, technological advancement, green SC management, and SBP in China. These outcomes guide regulators in making regulations related to sustainable performance using the best practice of SC, technology advancement, and knowledge management., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A series of density functional theory calculations were performed to understand the bonding and interaction of hydrogen adsorption on two-dimensional silicon carbide obtained from molecular dynamics simulation. The converged energy results pointed out that the H atom can sufficiently bond to 2D SiC at the top sites (atop Si and C), of which the most stable adsorption site is T Si . The vibrational properties along with the zero-point energy were incorporated into the energy calculations to further understand the phonon effect of the adsorbed H. Most of the 2D SiC structure deformations caused by the H atoms were found at the adsorbent atom along the vertical axis. For the first time, five SiC defect formations, including the quadrilateral-octagon linear defect (8-4), the silicon interstitial defect, the divacancy (4-10-4) defect, the divacancy (8-4-4-8) defect, and the divacancy (4-8-8-4) defect, were investigated and compared with previous 2D defect studies. The linear defect (8-4) has the lowest formation energy and is most likely to be formed for SiC materials. Furthermore, hydrogen atoms adsorb more readily on the defect surface than on the pristine SiC surface., Competing Interests: The authors declare no competing financial interest., (© 2022 The Authors. Published by American Chemical Society.)

The neurobiology of autism has been shown to involve alterations in cortical morphology and gamma-aminobutyric acid A (GABA A ) receptor density. We hypothesized that GABA A receptor binding potential (GABA A R BP ND ) would correlate with cortical thickness, but their correlations would differ between autistic adults and typically developing (TD) controls. We studied 50 adults (23 autism, 27 TD, mean age of 27 years) using magnetic resonance imaging to measure cortical thickness, and [ 18 F]flumazenil positron emission tomography imaging to measure GABA A R BP ND . We determined the correlations between cortical thickness and GABA A R BP ND by cortical lobe, region-of-interest, and diagnosis of autism spectrum disorder (ASD). We also explored potential sex differences in the relationship between cortical thickness and autism characteristics, as measured by autism spectrum quotient (AQ) scores. Comparing autism and TD groups, no significant differences were found in cortical thickness or GABA A R BP ND . In both autism and TD groups, a negative relationship between cortical thickness and GABA A R BP ND was observed in the frontal and occipital cortices, but no relationship was found in the temporal or limbic cortices. A positive correlation was seen in the parietal cortex that was only significant for the autism group. Interestingly, in an exploratory analysis, we found sex differences in the relationships between cortical thickness and GABA A R BP ND , and cortical thickness and AQ scores in the left postcentral gyrus. LAY SUMMARY: The thickness of the brain cortex and the density of the receptors associated with inhibitory neurotransmitter GABA have been hypothesized to underlie the neurobiology of autism. In this study, we found that these biomarkers correlate positively in the parietal cortex, but negatively in the frontal and occipital cortical regions of the brain. Furthermore, we collected preliminary evidence that the correlations between cortical thickness and GABA receptor density are sexdependent in a brain region where sensory inputs are registered., (© 2022 International Society for Autism Research and Wiley Periodicals LLC.)

The purpose of this policy statement is to update the 2004 American Academy of Pediatrics clinical report and provide enhanced guidance for institutions, administrators, and providers in the development and operation of a pediatric intermediate care unit (IMCU). Since 2004, there have been significant advances in pediatric medical, surgical, and critical care that have resulted in an evolution in the acuity and complexity of children potentially requiring IMCU admission. A group of 9 clinical experts in pediatric critical care, hospital medicine, intermediate care, and surgery developed a consensus on priority topics requiring updates, reviewed the relevant evidence, and, through a series of virtual meetings, developed the document. The intended audience of this policy statement is broad and includes pediatric critical care professionals, pediatric hospitalists, pediatric surgeons, other pediatric medical and surgical subspecialists, general pediatricians, nurses, social workers, care coordinators, hospital administrators, health care funders, and policymakers, primarily in resource-rich settings. Key priority topics were delineation of core principles for an IMCU, clarification of target populations, staffing recommendations, and payment., (Copyright © 2022 by the American Academy of Pediatrics.)

Water purification is becoming one of the most pertinent environmental issues throughout the world. Among common types of water pollution involving heavy metals, pharmaceutical drugs, textile dyes, personal care products, and other persistent organic pollutants, the pollution of antibiotic drugs is increasingly emerging due to their adverse effects on microorganisms, aquatic animals, and human health. Therefore, the treatment of such contaminants is very necessary to reduce the concentration of antibiotic pollutants to permissible levels prior to discharge. Herein, we report the use of NiFe 2 O 4 @C composites from a bimetallic-based metal-organic framework Ni-MIL-88B(Fe) for removal of ciprofloxacin (CFX) and tetracycline (TCC). The effect of production temperatures (600-900 °C), solution pH (2-10), NiFe 2 O 4 @C dose (0.05-0.2 g/L), concentration of antibiotics (10-60 mg/L), and uptake time (0-480 min) was investigated systematically. Response surface methodology and central composite design were applied for quadratic models to discover optimum conditions of antibiotic adsorption. With high coefficients of determination (R 2 = 0.9640-0.9713), the proposed models were significant statistically. Under proposed optimum conditions, the adsorption capacity for CFX and TCC were found at 256.244, and 105.38 mg/g, respectively. Recyclability study was employed and found that NiFe 2 O 4 @C-900 could be reused for up to three cycles, offering the potential of this composite as a good adsorbent for removal of emergent antibiotics.

In this work, the perturbation-induced phase transitions in noncentrosymmetric quantum spin Hall insulators (QSHIs) are analytically addressed. In particular, the dilute charged impurity, the electric field, and the Zeeman splitting field are considered within the tight-binding Hamiltonian model, Green's function approach, and the Born approximation. Following the C 3 v symmetry breaking in the PbBiI compound as a representative QSHI, the band gap becomes larger via the electric field, while the system transits to the semimetallic phase via the dilute charged impurities and Zeeman field, modifying the degenerate states in the electronic density of states. While the coexistence of electric field and impurities demonstrate that the system backs to its initial semiconducting phase, the combined Zeeman field and impurities do not alter the robust semimetallic phase., (© 2021 IOP Publishing Ltd.)

Alterations in sensorimotor functions are common in individuals with autism spectrum disorder (ASD). Such aberrations suggest the involvement of the thalamus due to its key role in modulating sensorimotor signaling in the cortex. Although previous research has linked atypical thalamocortical connectivity with ASD, investigations of this association in high-functioning adults with autism spectrum disorder (HFASD) are lacking. Here, for the first time, we investigated the resting-state functional connectivity of the thalamus, medial prefrontal, posterior cingulate, and left dorsolateral prefrontal cortices and its association with symptom severity in two matched cohorts of HFASD. The principal cohort consisted of 23 HFASD (mean[SD] 27.1[8.9] years, 39.1% female) and 20 age- and sex-matched typically developing controls (25.1[7.2] years, 30.0% female). The secondary cohort was a subset of the ABIDE database consisting of 58 HFASD (25.4[7.8] years, 37.9% female) and 51 typically developing controls (24.4[6.7] years, 39.2% female). Using seed-based connectivity analysis, between-group differences were revealed as hyperconnectivity in HFASD in the principal cohort between the right thalamus and bilateral precentral/postcentral gyri and between the right thalamus and the right superior parietal lobule. The former was associated with autism-spectrum quotient in a sex-specific manner, and was further validated in the secondary ABIDE cohort. Altogether, we present converging evidence for thalamocortical hyperconnectivity in HFASD that is associated with symptom severity. Our results fill an important knowledge gap regarding atypical thalamocortical connectivity in HFASD, previously only reported in younger cohorts.

Background: Little is known about the natural history of asymptomatic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection., Methods: We conducted a prospective study at a quarantine center for coronavirus disease 2019 in Ho Chi Minh City, Vietnam. We enrolled quarantined people with reverse-transcription polymerase chain reaction (RT-PCR)-confirmed SARS-CoV-2 infection, collecting clinical data, travel and contact history, and saliva at enrollment and daily nasopharyngeal/throat swabs (NTSs) for RT-PCR testing. We compared the natural history and transmission potential of asymptomatic and symptomatic individuals., Results: Between 10 March and 4 April 2020, 14 000 quarantined people were tested for SARS-CoV-2; 49 were positive. Of these, 30 participated in the study: 13 (43%) never had symptoms and 17 (57%) were symptomatic. Seventeen (57%) participants imported cases. Compared with symptomatic individuals, asymptomatic people were less likely to have detectable SARS-CoV-2 in NTS collected at enrollment (8/13 [62%] vs 17/17 [100%]; P = .02). SARS-CoV-2 RNA was detected in 20 of 27 (74%) available saliva samples (7 of 11 [64%] in the asymptomatic group and 13 of 16 [81%] in the symptomatic group; P = .56). Analysis of RT-PCR positivity probability showed that asymptomatic participants had faster viral clearance than symptomatic participants (P < .001 for difference over the first 19 days). This difference was most pronounced during the first week of follow-up. Two of the asymptomatic individuals appeared to transmit SARS-CoV-2 to 4 contacts., Conclusions: Asymptomatic SARS-CoV-2 infection is common and can be detected by analysis of saliva or NTSs. The NTS viral loads fall faster in asymptomatic individuals, but these individuals appear able to transmit the virus to others., (© The Author(s) 2020. Published by Oxford University Press for the Infectious Diseases Society of America.)

Background : COVID-19 is a respiratory disease caused by a novel coronavirus (SARS-CoV-2) and causes substantial morbidity and mortality. There is currently no vaccine to prevent COVID-19 or therapeutic agent to treat COVID-19. This clinical trial is designed to evaluate chloroquine as a potential therapeutic for the treatment of hospitalised people with COVID-19. We hypothesise that chloroquine slows viral replication in patients with COVID-19, attenuating the infection, and resulting in more rapid decline of viral load in throat/nose swabs. This viral attenuation should be associated with improved patient outcomes. Method : The study will start with a 10-patient prospective observational pilot study following the same entry and exclusion criteria as for the randomized trial and undergoing the same procedures. The main study is an open label, randomised, controlled trial with two parallel arms of standard of care (control arm) versus standard of care with 10 days of chloroquine (intervention arm) with a loading dose over the first 24 hours, followed by 300mg base orally once daily for nine days. The study will recruit patients in three sites in Ho Chi Minh City, Vietnam: the Hospital for Tropical Diseases, the Cu Chi Field Hospital, and the Can Gio COVID hospital. The primary endpoint is the time to viral clearance from throat/nose swab, defined as the time following randomization until the midpoint between the last positive and the first of the negative throat/nose swabs. Viral presence will be determined using RT-PCR to detect SARS-CoV-2 RNA. Discussion:  The results of the study will add to the evidence-based guidelines for management of COVID-19. Given the enormous experience of its use in malaria chemoprophylaxis, excellent safety and tolerability profile, and its very low cost, if proved effective then chloroquine would be a readily deployable and affordable treatment for patients with COVID-19. Trial registration: Clinicaltrials.gov NCT04328493 31/03/2020., Competing Interests: No competing interests were disclosed., (Copyright: © 2020 Kestelyn E et al.)

In this paper, the potential of engineering and manipulating the electronic heat capacity and Pauli susceptibility of pristine and perturbed hydrogenated AA-stacked graphene, SiC (silicon carbide), and h-BN (hexagonal boron nitride) bilayers is studied using a designed transverse Zeeman magnetic field and the dilute charged impurity. The tight-binding Hamiltonian model, the Born approximation and the Green's function method describe the carrier dynamics up to a certain degree. The unperturbed results show that the heat capacity and susceptibility of all bilayers increase with different hydrogenation doping configurations. We also found that the maximum heat capacity and susceptibility relates to the chair-like and table-like configurations. Also, the graphene possesses the highest activity compared to SiC and h-BN lattices due to its zero on-site energies. For the Zeeman magnetic field-induced Schottky anomaly and the Néel temperature corresponding to the maximum electronic heat capacity, EHC Max. , and Pauli spin paramagnetic susceptibility, PSPS Max. , respectively, the pristine EHC Max. (PSPS Max. ) decreases (increases) with the Zeeman field. On the other hand, the corresponding results for reduced table-like and reduced chair-like lattices illustrate that both EHC Max. and PSPS Max. decrease with the Zeeman field, on average. However, under the influence of the dilute charged impurity, the pristine EHC Max. of graphene (SiC and h-BN) decreases (increases) with impurity concentration for all configurations while the corresponding PSPS Max. fluctuates (decreases) for the pristine (reduced table-like and reduced chair-like) case. These findings introduce hydrogenated AA-stacked bilayers as versatile candidates for real applications., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Fertilizer is applied widely to improve the productivity of plantations. Traditionally, fertilization is conducted in spring and/or in the early rainy season, and it is believed to support the growth of planted trees in the growing season. Little attention to date has been paid on identification of the optimal timing of fertilization and fertilizer dose. In this study, application of the fine root monitoring technique in identifying optimal fertilization timing for an Acacia plantation in Vietnam is described. The study used two fertilizer doses (100 and 200 g NPK/tree) and three fertilization timings (in spring; in the early rainy season; and based on the fine root monitoring technique to identify when the fine roots reach their growth peak). As expected fertilization timings significantly affected growth and above-ground biomass (AGB) of the plantation. Fertilization based on the fine root monitoring technique resulted in the highest growths and AGB, followed by fertilization in the early rainy season and then in spring. Applying fertilizer at 200 g NPK/tree based on the fine root monitoring technique increased diameter at breast height (DBH) by 16%, stem height by 8%, crown diameter (Dc) by 16%, and AGB by 40% as compared to early rainy season fertilization. Increases of 32% DBH, 23% stem height, 44% Dc, and 87% AGB were found in fertilization based on fine root monitoring technique compared to spring fertilization. This study concluded that forest growers should use the fine root monitoring technique to identify optimal fertilization timing for higher productivity., Competing Interests: The authors have declared that no competing interests exist.

Bombyx mori fibroin has been widely used since a long time ago and has become a popular material. Here, we carry out a molecular dynamics simulation-based docking simulation of a small fragment of graphene in order to seek the best binding position on the N-termini domain of Bombyx mori fibroin. We report the best binding position, of which binding free energy falls at -54.8 kJ mol-1, indicating the strong binding. The further analysis of the binding pathway shows that this position is selective for single layered graphene rather than multi-layered graphene within our limited simulation times. Via comparing the RAMAN spectra of the corresponding binding pose of atomic clusters, we report the change in the bands compared with free standing graphene fragments, implying the change in molecular orbitals.

Introduction: Surgical resection of oesophageal cancer is a major procedure with potential for significant morbidity and mortality. Patient selection can be challenging, as operative benefit must be balanced against risk and impact on quality of life. This study defines modern trends in patient selection, and evaluates the impact of age, stage, and comorbidities on complications and survival following oesophagectomy, in a tertiary Australian experience., Methods: Data were compiled across two 15-year operative eras ('Era 1': 1981-1995; and 'Era 2': 1996-2010), with patients followed minimum 3 years. A total of 180 unselected records were analysed (powered for a relative hazard ratio of 0.5). Analyses defined patient selection trends, and for Era 2, the impact of age, comorbidities (Charlson score), and disease (T/N stage) on complications (Clavien-Dindo grade) and survival (Kaplan-Meier). A further sub-analysis was conducted with data divided into three 10-year periods., Results: The age of operated patients increased from Era 1 to 2 (mean+5 years; P<0.001), but survival and complication rates were unchanged, including in patients≥75 years (P>0.5). In Era 2, reflecting recent practice, survival duration matched T/N stage (P<0.001) but was independent of age at surgery (P=0.56) and comorbidity score (P=0.78). However, grade of worst post-operative complication, including death (rate: 3.8%), was correlated with both age (P<0.01) and comorbidity score (P<0.01)., Discussion: Older patients are now undergoing oesophagectomy. However, if they are selected appropriately, then older patients and those with comorbidities can expect similar stage-matched survival outcomes to younger fitter patients, despite their higher operative risk. Poor outcomes persist in patients with locally advanced disease, and selection in this group should prioritise quality of life.

Background: To examine the rate of Helicobacter pylori infection in a group of symptomatic patients using rapid urease test and antral histology and report on their treatment outcomes., Methods: From a single-surgeon series in a tertiary teaching hospital, 200 consecutive symptomatic patients undergoing their first diagnostic upper gastrointestinal endoscopy during 2003-2004 were included in this retrospective audit. Campylobacter-like organism (CLO) test results were compared with the results of conventional histological examination plus Giemsa staining. Eradication rates and demographic data were collected., Results: One hundred and seventy-nine of 200 (89.5%) patients had antral biopsies for histology and for CLOtest. The CLOtest was positive in 31% (56/179) of patients. Thirty-two of 56 (57%) patients had an early-positive CLOtest (<24 h). The remaining 43% (24/56) of patients had a late-positive CLOtest (>24 h). Histological examination of the antral biopsy showed evidence of H. pylori in 28/56 (50%) patients with a positive CLOtest. H. pylori eradication therapy was prescribed in 51/56 (91%) patients with a positive CLOtest. Forty-five of 51 (88%) of these patients had a subsequent urea breath test. Urea breath test was positive in 17/45 (29%) patients after H. pylori eradication therapy was prescribed., Conclusion: Rate of H. pylori in this series of symptomatic patients was 31%. As nearly half of the CLOtests became positive after 24 h, we suggest that the CLOtest should be re-examined 24 h after gastroscopy. Standard antral histology and CLOtest were in agreement in only 50% of cases, suggesting that the CLOtest is a more sensitive test. The failure rate of 29% for initial H. pylori eradication therapy reinforces the need for follow up after treatment.

Reconstitution of high-affinity agonist binding at the beta2-adrenoceptor (beta2AR) expressed in Sf9 insect cells requires a large excess of the stimulatory G-protein of adenylyl cyclase, Gsalpha, relative to receptor [R. Seifert, T. W. Lee, V. T. Lam & B. K. Kobilka, (1998) Eur. J. Biochem. 255, 369-382]. In a fusion protein of the beta2AR and Gsalpha (beta2AR-Gsalpha), which has only a 1 : 1 stoichiometry of receptor and G-protein, high-affinity agonist binding and agonist-stimulated GTP hydrolysis, guanosine 5'-O-(3-thiotriphosphate) (GTP[S]) binding and adenylyl cyclase (AC) activation are more efficient than in the nonfused coexpression system. In order to analyze the stability of the receptor/G-protein interaction, we constructed a fusion protein with a thrombin-cleavage site between beta2AR and Gsalpha (beta2AR-TS-Gsalpha). beta2AR-TS-Gsalpha efficiently reconstituted high-affinity agonist binding, agonist-stimulated GTP hydrolysis, GTP[S] binding and AC activation. Thrombin cleaves approximately 70% of beta2AR-TS-Gsalpha molecules in Sf9 membranes. Thrombin cleavage did not impair high-affinity agonist binding and GTP[S] binding but strongly reduced ligand-regulated GTPase activity and AC activity. We conclude that fusion of the beta2AR to Gsalpha promotes tight physical association of the two partners and that this association remains stable for a single activation/deactivation cycle even after cleavage of the link between the receptor and G-protein. Dilution of Gsalpha in the membrane and release of activated Gsalpha into the cytosol can both prevent cleaved beta2AR-TS-Gsalpha from undergoing multiple activation/deactivation cycles.

In most studies, coupling of the beta2-adrenoceptor (beta2AR) to the stimulatory, heterotrimeric GTP-binding protein of adenylyl cyclase the (Gs) is studied indirectly by measuring adenylyl cyclase activation. The aim of this study was to establish a model system in which beta2AR-Gs interactions can be studied directly at the level of the G-protein. We expressed the beta2AR alone, in combination with the alpha-subunit of Gs (G(s alpha)), and as fusion protein with G(s alpha) (beta2AR-G(s alpha)) in Sf9 insect cells. The beta2AR expressed alone couples poorly to the endogenous G(s alpha)-like G-protein of Sf9 cells since no high-affinity agonist binding could be detected, and the effects of agonist and inverse agonist on adenylyl cyclase, high-affinity GTPase and guanosine 5'-O-(3-thiotriphosphate) (GTP[S]) binding were small. Beta2AR-G(s alpha) reconstituted high-affinity agonist binding and regulated adenylyl cyclase more effectively than the beta2AR co-expressed with a large excess of G(s alpha). In membranes expressing beta2AR-G(s alpha), highly effective agonist- and inverse agonist regulation of high-affinity GTP hydrolysis and GTP[S] binding was observed. In contrast, agonist and inverse agonist regulation of GTP hydrolysis and GTP[S] binding in membranes expressing beta2AR and G(s alpha) as separate proteins was difficult to detect. Our data show that the beta2AR interacts with G(s alpha) more efficiently when expressed as a fusion protein than when expressed with an excess of non-fused G(s alpha). The beta2AR-G(s alpha) fusion protein provides a very sensitive model system to study the regulation of Gs function by beta2AR agonists and inverse agonists directly at the level of the G-protein.

An expression plasmid encoding the extracellular domain of the 75-kDa human tumor necrosis factor (TNF) type 2 receptor (TNF-R2) was constructed and used to generate a stable cell line secreting soluble TNF-R2 (sTNF-R2). Purified sTNF-R2 was resolved by SDS-PAGE into one band of approximate M(r) 43,000, consistent with a molecular weight of 36,000 +/- 4800 obtained by sedimentation equilibrium analysis. The apparent molecular weight observed by gel filtration chromatography was approximately 136,000. Glycosylation analysis revealed that Asn-149 is fully glycosylated, while Asn-171 is incompletely glycosylated (approximately 50%), and that a proline-, serine-, and threonine-rich region (residues 175-234) contains O-linked carbohydrate structures. Scatchard analysis of [125I]TNF-alpha and [125I]TNF-beta binding to sTNF-R2 gave dissociation constants (Kd) of 0.3 and 0.75 nM, respectively, comparable to those observed for intact cell-surface TNF-R2. The sTNF-R2 was found to block the cytotoxicity of both TNF-alpha and TNF-beta in a murine L-M cell assay. The sizes of the sTNF-R2.TNF-alpha and sTNF-R2.TNF-beta complexes determined by gel filtration chromatography were approximately 322 and 204 kDa, respectively. The stoichiometry of the sTNF-R2.TNF-alpha and sTNF-R2.TNF-beta complexes were examined by size-exclusion chromatography, sedimentation equilibrium, and cross-linking. The data from these studies suggest that at least two molecules of sTNF-R2 can bind to a single TNF-alpha or TNF-beta trimer.

A complete pathway for Azorhizobium caulinodans nicotinate catabolism has been determined from mutant phenotype analyses, isolation of metabolic intermediates, and structural studies. Nicotinate serves as a respiratory electron donor to O2 via a membrane-bound hydroxylase and a specific c-type cytochrome oxidase. The resulting oxidized product, 6-hydroxynicotinate, is next reduced to 1,4,5,6-tetrahydro-6-oxonicotinate. Hydrolytic ring breakage follows, with release of pyridine N as ammonium. Decarboxylation then releases the nicotinate C-7 carboxyl group as CO2, and the remaining C skeleton is then oxidized to yield glutarate. Transthioesterification with succinyl coenzyme A (succinyl-CoA) yields glutaryl-CoA, which is then oxidatively decarboxylated to yield crotonyl-CoA. As with general acyl beta oxidation, L-beta-hydroxybutyryl-CoA, acetoacetyl-CoA, and finally two molecules of acetyl-CoA are produced. In sum, nicotinate is catabolized to yield two CO2 molecules, two acetyl-CoA molecules, and ammonium. Nicotinate catabolism stimulates Azorhizobium N2 fixation rates in culture. Nicotinate catabolism mutants still able to liberate pyridine N as ammonium retain this capability, whereas mutants so blocked do not. From, mutant analyses and additional physiological tests, N2 fixation stimulation is indirect. In N-limited culture, nicotinate catabolism augments anabolic N pools and, as a consequence, yields N2-fixing cells with higher dinitrogenase content.

An expression plasmid encoding the human 75-kDa tumor necrosis factor (TNF) type 2 receptor (TNF-R2) was constructed and used to generate a stable human cell line (293/TNF-R2) overexpressing TNF-R2. Ligand binding analysis revealed high affinity binding (Kd = 0.2 nM) with approximately 94,000 +/- 7,500 sites/cell for 125I-TNF-alpha and approximately 5-fold lower affinity for TNF-beta (Kd = 1.1 nM) with 264,000 +/- 2,000 sites/cell. Cross-linking of 125I-TNF-alpha and 125I-TNF-beta to 293/TNF-R2 cells yielded predominant complexes with apparent molecular weights of 211,000 for TNF-alpha and 205,000 and 244,000 for TNF-beta, suggesting these complexes contain two or three TNF-R2 molecules. Immunoprecipitation of TNF-R2 from 32P-labeled 293/TNF-R2 cells demonstrated that the receptor is phosphorylated. The majority (97%) of 32Pi incorporation was found in serine residues with a very low level of incorporation (3%) in threonine residues. TNF-alpha treatment of 293/TNF-R2 cells did not significantly affect the degree or pattern of phosphorylation. Cell surface-bound 125I-TNF-alpha was slowly internalized by the 293/TNF-R2 cell line with a t1/2 = 25 min. Shedding of the extracellular domain of TNF-R2 was induced by 4 beta-phorbol 12-myristate 13-acetate but not by TNF-alpha or TNF-beta.