This study was conducted to examine a current baseline profile of antimicrobial resistance and virulence of Escherichia coli isolated from foods commonly sold in the market place in Vietnam. E. coli were isolated from 180 samples of raw meat, poultry and shellfish and also isolated from 43 chicken faeces samples. Ninety-nine E. coli isolates recovered from all sources were selected for the investigation of their susceptibility to 15 antimicrobial agents by the disk diffusion method. Eighty-four percent of the isolates were resistant to one or more antibiotics, and multi-resistance, defined as resistance to at least 3 different classes of antibiotics, was detected in all sources. The rates of multi-resistance were up to 89.5% in chicken, 95% in chicken faeces and 75% in pork isolates. Resistance was most frequently observed to tetracycline (77.8%), sulfafurazole (60.6%), ampicillin (50.5%), amoxicillin (50.5%), trimethoprim (51.5%), chloramphenicol (43.4%), streptomycin (39.4%), nalidixic acid (34.3%) and gentamicin (24.2%). In addition, the isolates also displayed resistance to fluoroquinolones (ciprofloxacin 16.2%, norfloxacin 17.2%, and enrofloxacin 21.2%), with chicken isolates showing the highest rates of resistance to these antibiotics (52.6-63.2%). Thirty-eight multi-resistant isolates were selected for further the examination of antibiotic resistance genes and were also evaluated for virulence gene profiles by multiplex and uniplex polymerase chain reaction. The beta-lactam TEM gene and tetracycline resistance tetA, tetB genes were frequently detected in the tested isolates (84.2% and 89.5% respectively). Genes which are responsible for resistance to streptomycin (aadA) (68.4%), chloramphenicol (cmlA) (42.1%), sulfonamides (sulI) (39.5%), trimethoprim (dhfrV) (26.3%) and kanamycin (aphA-1) (23.7%) were also widely distributed. Plasmid-mediated ampC genes were detected in E. coli isolates from chicken and pork. The isolates were tested for the presence of 58 virulence genes for adhesins, toxins, capsule synthesis, siderophores, invasins and others from different E. coli pathotypes. All of the tested isolates contained at least one virulence gene and there were 16 genes detected. Virulence genes detected were fimH (92.1%), bmaE (84.2%), TSPE4.C2 (42.1%), aidA AIDA-I (orfB) (31.6%), east1 (26.3%), traT (23.7%), and others including fyuA, iutA, chuA, yjaA, iss, iroN(E. coli), ibeA, aah (orfA), iha and papG allele III (10.5-2.6%). Typical toxin genes produced by enterohemorrhagic and enterotoxigenic E. coli pathotypes (a heat-stable toxin (ST), heat-labile toxin (LT) and Shiga toxin stx1, stx2) were not detected in any of these 38 isolates. The study has revealed that E. coli in raw foods is a significant reservoir of resistance and virulence genes.

Adaptive immune responses are characterized by substantial restructuring of secondary lymphoid organs. The molecular and cellular factors responsible for virus-induced lymphoid remodeling are not well known to date. Here we applied optical projection tomography, a mesoscopic imaging technique, for a global analysis of the entire 3-dimensional structure of mouse peripheral lymph nodes (PLNs), focusing on B-cell areas and high endothelial venule (HEV) networks. Structural homeostasis of PLNs was characterized by a strict correlation between total PLN volume, B-cell volume, B-cell follicle number, and HEV length. After infection with lymphocytic choriomeningitis virus, we observed a substantial, lymphotoxin (LT) beta-receptor-dependent reorganization of the PLN microarchitecture, in which an initial B-cell influx was followed by 3-fold increases in PLN volume and HEV network length on day 8 after infection. Adoptive transfer experiments revealed that virus-induced PLN and HEV network remodeling required LTalpha(1)beta(2)-expressing B cells, whereas the inhibition of vascular endothelial growth factor-A signaling pathways had no significant effect on PLN expansion. In summary, lymphocytic choriomeningitis virus-induced PLN growth depends on a vascular endothelial growth factor-A-independent, LT- and B cell-dependent morphogenic pathway, as revealed by an in-depth mesoscopic analysis of the global PLN structure.

Loss of the JunB/AP-1 transcription factor induces a myeloproliferative disease (MPD) arising from the hematopoietic stem cell (HSC) compartment. Here, we show that junB inactivation deregulates the cell-cycle machinery and increases the proliferation of long-term repopulating HSCs (LT-HSCs) without impairing their self-renewal or regenerative potential in vivo. We found that JunB loss destabilizes a complex network of genes and pathways that normally limit myeloid differentiation, leading to impaired responsiveness to both Notch and TGF-beta signaling due in part to transcriptional deregulation of the Hes1 gene. These results demonstrate that LT-HSC proliferation and differentiation are uncoupled from self-renewal and establish some of the mechanisms by which JunB normally limits the production of myeloid progenitors, hence preventing initiation of myeloid malignancies.

OBJECTIVE

Exosomes isolated from the plasma of newly diagnosed acute myeloid leukemia (AML) patients have elevated protein and transforming growth factor-beta 1 (TGF-β1) contents and inhibit natural killer (NK) cell cytotoxicity (Haematologica 96, p. 1302, 2011). A potential role of exosomes in predicting responses to chemotherapy (CT) was evaluated in AML patients undergoing treatment.

METHODS

Plasma was obtained from AML patients at diagnosis (n = 16); post-induction CT (n = 9); during consolidation CT (n = 10); in long-term remission (Lt-CR, n = 5); and from healthy volunteers (n = 7). Exosomes were isolated by size-exclusion chromatography and ultracentrifugation. The exosomal protein, soluble TGFβ-1 levels (ELISA), and the TGF-β1 profiles (western blots) were compared among patients' cohorts. The results were correlated with the patients' cytogenetic profile, percentage of leukemic blast, and outcome.

RESULTS

At diagnosis, protein and TGF-β1 levels were higher (p < 0.009 and p < 0.004) in AML than control exosomes. These values decreased after induction CT (p < 0.05 and p < 0.004), increased during consolidation CT (p < 0.02 and p < 0.005), and normalized in Lt-CR. While TGF-β1 and protein levels tracked one another, TGF-β1 pro-peptide, latency-associated peptide (LAP), or mature TGF-β1 differentially decorated exosomes isolated before, during, and after CT. Only TGF-β1 pro-peptide was seen in exosomes of controls or Lt-CR patients. During consolidation CT, exosomes carried TGF-β1 pro-peptide, LAP, and low levels of mature TGF-β1. NK cell co-incubation with AML exosomes carrying all three TGF-β1 forms induced down-regulation of NKG2D expression.

CONCLUSIONS

Changes in exosomal protein and/or TGF-β1 content may reflect responses to CT. The exosomal profile may suggest the presence of residual disease in patients considered to have achieved complete remission.

<A<em>b</em>stractText>Despite improvements in mu<em>lt</em>idisciplinary management, patients with <em>b</em>iliary tract cancer have a poor outcome. Only 20% of patients are eligi<em>b</em>le for surgical resection with curative intent, with 5-year overall survival of less than 10% for all patients. To our knowledge, no studies have descri<em>b</em>ed a <em>b</em>enefit of adjuvant therapy. We aimed to determine whether adjuvant capecita<em>b</em>ine improved overall survival compared with o<em>b</em>servation following surgery for <em>b</em>iliary tract cancer.</A<em>b</em>stractText><p><div>(<em>b</em>)METHODS</<em>b</em>)</div>This randomised, controlled, mu<em>lt</em>icentre, phase 3 study was done across 44 specialist hepatopancreato<em>b</em>iliary centres in the UK. Eligi<em>b</em>le patients were aged 18 years or older and had histologically confirmed cholangiocarcinoma or muscle-invasive gall<em>b</em>ladder cancer who had undergone a macroscopically complete resection (which includes liver resection, pancreatic resection, or, less commonly, <em>b</em>oth) with curative intent, and an Eastern Cooperative Oncology Group performance status of less than 2. Patients who had not completely recovered from previous surgery or who had previous chemotherapy or radiotherapy for <em>b</em>iliary tract cancer were also excluded. Patients were randomly assigned 1:1 to receive oral capecita<em>b</em>ine (1250 mg/m² twice daily on days 1-14 of a 21-day cycle, for eight cycles) or o<em>b</em>servation commencing within 16 weeks of surgery. Treatment was not masked, and allocation concealment was achieved with a computerised minimisation algorithm that stratified patients <em>b</em>y surgical centre, site of disease, resection status, and performance status. The primary outcome was overall survival. As prespecified, analyses were done <em>b</em>y intention to treat and per protocol. This study is registered with EudraCT, num<em>b</em>er 2005-003318-13.</p><A<em>b</em>stractText>Between March 15, 2006, and Dec 4, 2014, 447 patients were enrolled; 223 patients with <em>b</em>iliary tract cancer resected with curative intent were randomly assigned to the capecita<em>b</em>ine group and 224 to the o<em>b</em>servation group. The data cutoff for this analysis was March 6, 2017. The median follow-up for all patients was 60 months (IQR 37-60). In the intention-to-treat analysis, median overall survival was 51·1 months (95% CI 34·6-59·1) in the capecita<em>b</em>ine group compared with 36·4 months (29·7-44·5) in the o<em>b</em>servation group (adjusted hazard ratio [HR] 0·81, 95% CI 0·63-1·04; p=0·097). In a protocol-specified sensitivity analysis, adjusting for minimisation factors and nodal status, grade, and gender, the overall survival HR was 0·71 (95% CI 0·55-0·92; p=0·010). In the prespecified per-protocol analysis (210 patients in the capecita<em>b</em>ine group and 220 in the o<em>b</em>servation group), median overall survival was 53 months (95% CI 40 to not reached) in the capecita<em>b</em>ine group and 36 months (30-44) in the o<em>b</em>servation group (adjusted HR 0·75, 95% CI 0·58-0·97; p=0·028). In the intention-to-treat analysis, median recurrence-free survival was 24·4 months (95% CI 18·6-35·9) in the capecita<em>b</em>ine group and 17·5 months (12·0-23·8) in the o<em>b</em>servation group. In the per-protocol analysis, median recurrence-free survival was 25·9 months (95% CI 19·8-46·3) in the capecita<em>b</em>ine group and 17·4 months (12·0-23·7) in the o<em>b</em>servation group. Adverse events were measured in the capecita<em>b</em>ine group only, and of the 213 patients who received at least one cycle, 94 (44%) had at least one grade 3 toxicity, the most frequent of which were hand-foot syndrome in 43 (20%) patients, diarrhoea in 16 (8%) patients, and fatigue in 16 (8%) patients. One (&<em>lt</em>;1%) patient had grade 4 cardiac ischaemia or infarction. Serious adverse events were o<em>b</em>served in 47 (21%) of 223 patients in the capecita<em>b</em>ine group and 22 (10%) of 224 patients in the o<em>b</em>servation group. No deaths were deemed to <em>b</em>e treatment related.</A<em>b</em>stractText><A<em>b</em>stractText>A<em>lt</em>hough this study did not meet its primary endpoint of improving overall survival in the intention-to-treat population, the prespecified sensitivity and per-protocol analyses suggest that capecita<em>b</em>ine can improve overall survival in patients with resected <em>b</em>iliary tract cancer when used as adjuvant chemotherapy following surgery and could <em>b</em>e considered as standard of care. Furthermore, the safety profile is managea<em>b</em>le, supporting the use of capecita<em>b</em>ine in this setting.</A<em>b</em>stractText><A<em>b</em>stractText>Cancer Research UK and Roche.</A<em>b</em>stractText>

Environmental and nontoxigenic strains of Vibrio cholerae 0-1 were examined for genes homologous to genes encoding Escherichia coli heat-labile enterotoxin (LT). Restriction fragments encoding LT A and B subunits were isolated from the recombinant plasmid EWD299 and labeled in vitro with 32P. These probes were then hybridized to deoxyribonucleic acid extracted from strains of V. cholerae and visualized by autoradiography. None of the nontoxigenic strains of V. cholerae 0-1 from Louisiana, Alabama, Maryland, Guam, Brazil, Bangladesh, or Great Britain hybridized with the LT probes, whereas all toxigenic strains exhibited homology. In addition, strains of V. cholerae non-0-1, "group F" vibrios, V. vulnificus, and Aeromonas hydrophila were tested, and all were negative except two strains of V. cholerae non-0-1. The presence of plasmids did not correlate with toxigenicity or nontoxigenicity in any of the species examined. Thus, it appears that these strains are not simple nontoxigenic mutants, but rather do not possess any genetic material encoding cholera toxin. Such strains therefore cannot revert and serve as a reservoir of cholera.

Hepatitis C virus (HCV) core protein is a multifunctional protein. We examined whether it can interact with cellular proteins, thus contributing to viral pathogenesis. Using the HCV core protein as a bait to screen a human liver cDNA library in a yeast two-hybrid screening system, we have isolated several positive clones encoding cellular proteins that interact with the HCV core protein. Interestingly, more than half of these clones encode the cytoplasmic domain of lymphotoxin-beta receptor (LT betaR), which is a member of the tumor necrosis factor receptor family. Their binding was confirmed by in vitro glutathione S-transferase fusion protein binding assay and protein-protein blotting assay to be direct and specific. The binding sites were mapped within a 58-amino-acid region of the cytoplasmic tail of LT betaR. The binding site in the HCV core protein was localized within amino acid residues 36 to 91 from the N terminus, corresponding to the hydrophilic region of the protein. In mammalian cells, the core protein was found to be associated with the membrane-bound LT betaR. Since the LT betaR is involved in germinal center formation and developmental regulation of peripheral lymphoid organs, lymph node development, and apoptotic signaling, the binding of HCV core protein to LT betaR suggests the possibility that this viral protein has an immunomodulating function and may explain the mechanism of viral persistence and pathogenesis of HCV.

It has been proposed that extracellular ATP may be involved in visceral mechanosensory transduction by activating ligand-gated ion channels (P2X receptors). In this study, we have investigated the effects of the P2X(3) agonist alpha,beta-methylene ATP (alpha,beta-meATP) and antagonist 2',3'-O-trinitrophenyl-ATP (TNP-ATP) on pelvic afferents innervating the urinary bladder using an in vitro mouse bladder-pelvic nerve preparation. Intravesical application of alpha,beta-meATP (0.03-1 mM) increased multifibre discharges in a concentration-dependent manner. The agonist potentiated, whereas TNP-ATP (0.03 mM) attenuated, the multifibre responses to bladder distensions. Single-unit analysis revealed that both high threshold (HT) fibres >> 15 mmHg; known to be associated with nociception) and low threshold (LT) fibres (< 15 mmHg; probably associated with non-nociceptive events) could be induced to discharge by intravesical alpha,beta-meATP (1 mM, 0.1 ml). The response of the vast majority (21/22, 95.5 %) of HT fibres to bladder distensions was enhanced with a significantly reduced threshold and an increased peak response after exposure to the agonist. On the other hand, 59.7 % (46/77) of LT fibres showed a greater peak and a slightly reduced threshold for response to bladder distension in the presence of alpha,beta-meATP. An additional 11 'silent' fibres became mechanosensitive after exposure to alpha,beta-meATP. TNP-ATP (0.03 mM) did not affect the threshold of LT fibres, but it reduced the peak response of some (22/51, 43.1 %) LT fibres. Conversely, the antagonist resulted in a markedly elevated threshold and reduced peak activity in the majority (13/16, 81.3 %) of HT fibres. The results support the view that P2X(3) receptor-mediated mechanisms contribute to both nociceptive and non-nociceptive (physiological) mechanosensory transduction in the urinary bladder.

<A<em>b</em>stractText>Preoperative chemoradiotherapy may improve the radical resection rate for resecta<em>b</em>le or <em>b</em>orderline resecta<em>b</em>le pancreatic cancer, <em>b</em>ut the overall <em>b</em>enefit is unproven.</A<em>b</em>stractText><A<em>b</em>stractText>In this randomized phase III trial in 16 centers, patients with resecta<em>b</em>le or <em>b</em>orderline resecta<em>b</em>le pancreatic cancer were randomly assigned to receive preoperative chemoradiotherapy, which consisted of 3 courses of gemcita<em>b</em>ine, the second com<em>b</em>ined with 15 × 2.4 Gy radiotherapy, followed <em>b</em>y surgery and 4 courses of adjuvant gemcita<em>b</em>ine or to immediate surgery and 6 courses of adjuvant gemcita<em>b</em>ine. The primary end point was overall survival <em>b</em>y intention to treat.</A<em>b</em>stractText><p><div>(<em>b</em>)RESULTS</<em>b</em>)</div>Between April 2013 and July 2017, 246 eligi<em>b</em>le patients were randomly assigned; 119 were assigned to preoperative chemoradiotherapy and 127 to immediate surgery. Median overall survival <em>b</em>y intention to treat was 16.0 months with preoperative chemoradiotherapy and 14.3 months with immediate surgery (hazard ratio, 0.78; 95% CI, 0.58 to 1.05; <i>P</i> = .096). The resection rate was 61% and 72% (<i>P</i> = .058). The R0 resection rate was 71% (51 of 72) in patients who received preoperative chemoradiotherapy and 40% (37 of 92) in patients assigned to immediate surgery (<i>P</i> &<em>lt</em>; .001). Preoperative chemoradiotherapy was associated with significantly <em>b</em>etter disease-free survival and locoregional failure-free interval as well as with significantly lower rates of pathologic lymph nodes, perineural invasion, and venous invasion. Survival analysis of patients who underwent tumor resection and started adjuvant chemotherapy showed improved survival with preoperative chemoradiotherapy (35.2 <i>v</i> 19.8 months; <i>P = .</i>029). The proportion of patients who suffered serious adverse events was 52% versus 41% (<i>P = .</i>096).</p><A<em>b</em>stractText>Preoperative chemoradiotherapy for resecta<em>b</em>le or <em>b</em>orderline resecta<em>b</em>le pancreatic cancer did not show a significant overall survival <em>b</em>enefit. A<em>lt</em>hough the outcomes of the secondary end points and predefined su<em>b</em>group analyses suggest an advantage of the neoadjuvant approach, additional evidence is required.</A<em>b</em>stractText>

To compare the function of the tumor necrosis factor (TNF) and lymphotoxin (LT)alpha/beta systems in the mature immune system, these two pathways were blocked with soluble receptor-immunoglobulin (R-Ig) fusion proteins in normal adult mice. Inhibition of LT alpha/beta signaling using LT betaR-Ig or a blocking monoclonal antibody against murine LT beta had profound effects. The spleen lacked discrete B cell follicles and the marginal zone was altered. Less marked changes were detected in lymph nodes. LT alpha/beta inhibition also prevented germinal center formation in the spleen and impaired Ig production in response to sheep red blood cells (SRBC) immunization. These results show that the LT alpha/beta system is required for the maintenance of splenic architecture and normal immune responses, and not simply for the development of peripheral immune organs during ontogeny. In contrast, inhibition of the TNF/LT alpha pathway with TNF-R55-Ig did not affect the splenic architecture or the anti-SRBC response. Splenic defects and impaired antibody responses are seen in TNF-deficient mice, suggesting that TNF is important during development. Therefore relative to TNF, the LT system has the dominant influence on splenic organization and anti-SRBC Ig formation in the adult mouse.

Several factors have been proposed to explain the high death rate of the coronavirus disease 2019 (COVID-19) outbreak, including hypertension and hypertension-related treatment with Renin Angiotensin System inhibitors. Also, age and multimorbidity might be confounders. No sufficient data are available to demonstrate their independent role. We designed a cross-sectional, observational, multicenter, nationwide survey in Italy to verify whether renin-angiotensin system inhibitors are related to COVID-19 severe outcomes. We analyzed information from Italian patients diagnosed with COVID-19, admitted in 26 hospitals. One thousand five hundred ninety-one charts (male, 64.1%; 66±0.4 years) were recorded. At least 1 preexisting condition was observed in 73.4% of patients, with hypertension being the most represented (54.9%). One hundred eighty-eight deaths were recorded (11.8%; mean age, 79.6±0.9 years). In nonsurvivors, older age, hypertension, diabetes mellitus, chronic obstructive pulmonary disease, chronic kidney disease, coronary artery diseases, and heart failure were more represented than in survivors. The Charlson Comorbidity Index was significantly higher in nonsurvivors compared with survivors (4.3±0.15 versus 2.6±0.05; P&lt;0.001). ACE (angiotensin-converting enzyme) inhibitors, diuretics, and β-blockers were more frequently used in nonsurvivors than in survivors. After correction by multivariate analysis, only age (P=0.0001), diabetes mellitus (P=0.004), chronic obstructive pulmonary disease (P=0.011), and chronic kidney disease (P=0.004) but not hypertension predicted mortality. Charlson Comorbidity Index, which cumulates age and comorbidities, predicts mortality with an exponential increase in the odds ratio by each point of score. In the COVID-19 outbreak, mortality is predicted by age and the presence of comorbidities. Our data do not support a significant interference of hypertension and antihypertensive therapy on COVID-19 lethality. Registration- URL: https://www.clinicaltrials.gov; Unique identifier: NCT04331574.

Keywords: COVID-19; Italy; hypertension; multimorbidity; odds ratio.

Importance: There are limitations in current diagnostic testing approaches for Alzheimer disease (AD).

Objective: To examine plasma tau phosphorylated at threonine 217 (P-tau217) as a diagnostic biomarker for AD.

Design, setting, and participants: Three cross-sectional cohorts: an Arizona-based neuropathology cohort (cohort 1), including 34 participants with AD and 47 without AD (dates of enrollment, May 2007-January 2019); the Swedish BioFINDER-2 cohort (cohort 2), including cognitively unimpaired participants (n = 301) and clinically diagnosed patients with mild cognitive impairment (MCI) (n = 178), AD dementia (n = 121), and other neurodegenerative diseases (n = 99) (April 2017-September 2019); and a Colombian autosomal-dominant AD kindred (cohort 3), including 365 PSEN1 E280A mutation carriers and 257 mutation noncarriers (December 2013-February 2017).

Exposures: Plasma P-tau217.

Main outcomes and measures: Primary outcome was the discriminative accuracy of plasma P-tau217 for AD (clinical or neuropathological diagnosis). Secondary outcome was the association with tau pathology (determined using neuropathology or positron emission tomography [PET]).

Results: Mean age was 83.5 (SD, 8.5) years in cohort 1, 69.1 (SD, 10.3) years in cohort 2, and 35.8 (SD, 10.7) years in cohort 3; 38% were women in cohort 1, 51% in cohort 2, and 57% in cohort 3. In cohort 1, antemortem plasma P-tau217 differentiated neuropathologically defined AD from non-AD (area under the curve [AUC], 0.89 [95% CI, 0.81-0.97]) with significantly higher accuracy than plasma P-tau181 and neurofilament light chain (NfL) (AUC range, 0.50-0.72; P &lt; .05). The discriminative accuracy of plasma P-tau217 in cohort 2 for clinical AD dementia vs other neurodegenerative diseases (AUC, 0.96 [95% CI, 0.93-0.98]) was significantly higher than plasma P-tau181, plasma NfL, and MRI measures (AUC range, 0.50-0.81; P &lt; .001) but not significantly different compared with cerebrospinal fluid (CSF) P-tau217, CSF P-tau181, and tau-PET (AUC range, 0.90-0.99; P > .15). In cohort 3, plasma P-tau217 levels were significantly greater among PSEN1 mutation carriers, compared with noncarriers, from approximately 25 years and older, which is 20 years prior to estimated onset of MCI among mutation carriers. Plasma P-tau217 levels correlated with tau tangles in participants with (Spearman ρ = 0.64; P &lt; .001), but not without (Spearman ρ = 0.15; P = .33), β-amyloid plaques in cohort 1. In cohort 2, plasma P-tau217 discriminated abnormal vs normal tau-PET scans (AUC, 0.93 [95% CI, 0.91-0.96]) with significantly higher accuracy than plasma P-tau181, plasma NfL, CSF P-tau181, CSF Aββlt; .05), but its performance was not significantly different compared with CSF P-tau217 (AUC, 0.96; P = .22).

Conclusions and relevance: Among 1402 participants from 3 selected cohorts, plasma P-tau217 discriminated AD from other neurodegenerative diseases, with significantly higher accuracy than established plasma- and MRI-based biomarkers, and its performance was not significantly different from key CSF- or PET-based measures. Further research is needed to optimize the assay, validate the findings in unselected and diverse populations, and determine its potential role in clinical care.

The recent development of quinoline-<em>b</em>ased PET tracers that act as fi<em>b</em>ro<em>b</em>last-activation-protein inhi<em>b</em>itors (FAPIs) demonstrated promising preclinical and clinical resu<em>lt</em>s. FAP is overexpressed <em>b</em>y cancer-associated fi<em>b</em>ro<em>b</em>lasts of several tumor entities. Here, we quantify the tumor uptake on ⁶⁸Ga-FAPI PET/CT of various primary and metastatic tumors to identify the most promising indications for future application. (<em>b</em>)Methods:</<em>b</em>)⁶⁸Ga-FAPI PET/CT scans were requested <em>b</em>y various referring physicians according to individual clinical indications that were considered insufficiently covered <em>b</em>y ¹⁸F-FDG PET/CT or other imaging modalities. All PET/CT was performed 1 h after injection of 122-312 MBq of ⁶⁸Ga-FAPI-04. We retrospectively identified 80 patients with histopathologically proven primary tumors or metastases or radiologically unequivocal metastatic lesions of histologically proven primary tumors. Tumor uptake was quantified <em>b</em>y SUV<su<em>b</em>)max</su<em>b</em>) and SUV<su<em>b</em>)mean</su<em>b</em>) (60% isocontour). (<em>b</em>)Resu<em>lt</em>s:</<em>b</em>) Eighty patients with 28 different tumor entities (54 primary tumors and 229 metastases) were evaluated. The highest average SUV<su<em>b</em>)max</su<em>b</em>) (>12) was found in sarcoma, esophageal, <em>b</em>reast, cholangiocarcinoma, and lung cancer. The lowest ⁶⁸Ga-FAPI uptake (average SUV<su<em>b</em>)max</su<em>b</em>) &<em>lt</em>; 6) was o<em>b</em>served in pheochromocytoma, renal cell, differentiated thyroid, adenoid cystic, and gastric cancer. The average SUV<su<em>b</em>)max</su<em>b</em>) of hepatocellular, colorectal, head-neck, ovarian, pancreatic, and prostate cancer was intermediate (SUV 6-12). SUV varied across and within all tumor entities. Because of low <em>b</em>ackground in muscle and <em>b</em>lood pool (SUV<su<em>b</em>)max</su<em>b</em>) &<em>lt</em>; 2), the tumor-to-<em>b</em>ackground contrast ratios were more than 3-fold in the intermediate and more than 6-fold in the high-intensity uptake group. (<em>b</em>)Conclusion:</<em>b</em>) Several highly prevalent cancers presented with remarka<em>b</em>ly high uptake and image contrast on ⁶⁸Ga-FAPI PET/CT. The high and rather selective tumor uptake may open up new applications for noninvasive tumor characterization, staging examinations, or radioligand therapy.

Immunodiffusion and biological neutralization studies demonstrated that the heat-labile enterotoxin (LT) from Escherichia coli has antigenic determinants in common with each of the isolated subunits (A and B) of the enterotoxin (choleragen) from Vibrio cholerae. Each of the enterotoxins also possesses unique antigenic specificities. Monospecific antiserum to LT was prepared by immunization with antigens derived by immune precipitation of E. coli cell-free supernatant with isolated specific anticholeragenoid antibodies. This antiserum neutralized the biological acitivity of both LT and cholera enterotoxin and recognized antigens of both in immunodiffusion. This antiserum was adsorbed with choleragenoid to remove antibodies directed against the shared "B" immunological determinants. The neutralizing effect of the antiserum on cholera toxin was completely removed, but the neutralizing activity against the E. coli preparations was retained, although somewhat reduced. Antisera to the isolated subunits (A and B) of cholera enterotoxin neutralized the biological activity of cholera enterotoxin and LT. These antisera also recognized the homologous and heterologous antigens in immunodiffusion. Multiple forms or conformations of LT and its components may explain the diversity of the properties which have been reported for it.

Mucosal immunisation may be used both to protect the mucosal surfaces against infections and as a means for immunological treatment of peripheral immunopathological disorders through the induction of systemic antigen-specific tolerance ('oral tolerance'). The development of mucosal vaccines, whether for prevention of infectious diseases or for oral tolerance immunotherapy, requires efficient antigen delivery and adjuvant systems that can help to present the appropriate vaccine or immunotherapy antigens to the mucosal immune system. The most potent (but also toxic) mucosal adjuvants are cholera toxin (CT) and the closely related Escherichia coli heat-labile enterotoxin (LT), and much effort and significant progress have been made recently to generate toxicologically acceptable derivatives of these toxins with retained adjuvant activity. Among these are the non-toxic, recombinantly produced cholera toxin B-subunit (CTB). CTB is a specific protective antigen component of a widely registered oral cholera vaccine as well as a promising vector for either giving rise to mucosal anti-infective immunity or for inducing peripheral anti-inflammatory tolerance to chemically or genetically linked foreign antigens administered mucosally. CT and CTB have also recently been used as combined vectors and adjuvants for markedly promoting ex vivo dendritic cell (DC) vaccination with different antigens and also steering the immune response to the in vivo-reinfused DCs towards either broad Th1 + Th2 + CTL immunity (CT) or Th2 or tolerance (CTB). Another type of mucosal adjuvants is represented by bacterial DNA or synthetic oligodeoxynucleotides containing CpG-motifs, which especially when linked to CTB have been found to effectively stimulate both innate and adaptive mucosal immune responses. The properties and clinical potential of these different classes of adjuvants are being discussed.

The thymus gives rise to two T cell lineages, alphabeta and gammadelta, that are thought to develop independently of one another. Hence, double positive (DP) thymocytes expressing CD4 and CD8 coreceptors are usually viewed simply as progenitors of CD4+ and CD8+ alphabeta T cells. Instead we report that DP cells regulate the differentiation of early thymocyte progenitors and gammadelta cells, by a mechanism dependent on the transcription factor RORgt, and the lymphotoxin (LT) beta receptor (LTbetaR). This finding provokes a revised view of the thymus, in which lymphoid tissue induction-type processes coordinate the developmental and functional integration of the two T cell lineages.

We have investigated the role of type I IFNs (IFN-alpha and -beta) in human T cell differentiation using anti-CD3 mAb and allogeneic, in vitro-derived dendritic cells (DC) as APCs. DC were very efficient activators of naive CD4+ T cells, providing necessary costimulation and soluble factors to support Th1 differentiation and expansion. Addition of IFN-alphabeta to DC/T cell cultures resulted in induction of T cell IL-10 production and inhibition of IFN-gamma, TNF-alpha, and LT secretion. Diminished T cell IFN-gamma production correlated with IFN-alphabeta-mediated inhibition of the p40 chain of the IL-12 heterodimer secreted by DC. Suppression of p40 IL-12 and IFN-gamma was not due to increased levels of IL-10 in these cultures, and production of IFN-gamma could be restored by exogenous IL-12. These data indicate that type I IFNs inhibit DC p40 IL-12 expression, which is required for development of IFN-gamma-producing CD4+ T cells. Furthermore, when T cells were restimulated without IFN-beta, these cells induced less p40 IL-12 from DC, suggesting that the functional properties of T cells may regulate DC function. Thus, IFN-alphabeta inhibits both IL-12-dependent and independent Th1 cytokine production and provides a mechanism for inhibition of IL-12-mediated immunity in viral infections.

(<em>b</em>)OBJECTIVE.</<em>b</em>) Confronting the new coronavirus infection known as coronavirus disease 2019 (COVID-19) is challenging and requires excluding patients with suspected COVID-19 who actually have other diseases. The purpose of this study was to assess the clinical features and CT manifestations of COVID-19 <em>b</em>y comparing patients with COVID-19 pneumonia with patients with non-COVID-19 pneumonia who presented at a fever o<em>b</em>servation department in Shanghai, China. (<em>b</em>)MATERIALS AND METHODS.</<em>b</em>) Patients were retrospectively enrolled in the study from January 19 through Fe<em>b</em>ruary 6, 2020. All patients underwent real-time reverse transcription-polymerase chain reaction (RT-PCR) testing. (<em>b</em>)RESULTS.</<em>b</em>) Eleven patients had RT-PCR test resu<em>lt</em>s that were positive for severe acute respiratory syndrome coronavirus 2, whereas 22 patients had negative resu<em>lt</em>s. No statistical difference in clinical features was o<em>b</em>served (<i>p</i> > 0.05), with the exception of leukocyte and platelet counts (<i>p</i> &<em>lt</em>; 0.05). The mean (± SD) interval <em>b</em>etween onset of symptoms and admission to the fever o<em>b</em>servation department was 4.40 ± 2.00 and 5.52 ± 4.00 days for patients with positive and negative RT-PCR test resu<em>lt</em>s, respectively. The frequency of opacifications in patients with positive resu<em>lt</em>s and patients with negative resu<em>lt</em>s, respectively, was as follows: ground-glass opacities (GGOs), 100.0% versus 90.9%; mixed GGO, 63.6% versus 72.7%; and consolidation, 54.5% versus 77.3%. In patients with positive RT-PCR resu<em>lt</em>s, GGOs were the most commonly o<em>b</em>served opacification (seen in 100.0% of patients) and were predominantly located in the peripheral zone (100.0% of patients), compared with patients with negative resu<em>lt</em>s (31.8%) (<i>p</i> = 0.05). The median num<em>b</em>er of affected lung lo<em>b</em>es and segments was higher in patients with positive RT-PCR resu<em>lt</em>s than in those with negative RT-PCR resu<em>lt</em>s (five vs 3.5 affected lo<em>b</em>es and 15 vs nine affected segments; <i>p</i> &<em>lt</em>; 0.05). A<em>lt</em>hough the air <em>b</em>ronchogram reticular pattern was more frequently seen in patients with positive resu<em>lt</em>s, centrilo<em>b</em>ular nodules were less frequently seen in patients with positive resu<em>lt</em>s. (<em>b</em>)CONCLUSION.</<em>b</em>) At the point during the COVID-19 out<em>b</em>reak when this study was performed, imaging patterns of mu<em>lt</em>ifocal, peripheral, pure GGO, mixed GGO, or consolidation with slight predominance in the lower lung and findings of more extensive GGO than consolidation on chest CT scans o<em>b</em>tained during the first week of illness were considered findings highly suspicious of COVID-19.

BACKGROUND

The role of eicosanoids, including leukotrienes (LTs) and prostaglandins (PGs), in chronic obstructive pulmonary disease (COPD) is uncertain. The aim of this study was to investigate whether eicosanoids are measurable in exhaled breath condensate (EBC), a non-invasive method of collecting airway secretions, in patients with stable mild to moderate COPD, and to show possible differences in their concentrations compared with control subjects.

METHODS

LTB(4), LTE(4), PGE(2), PGD(2)-methoxime, PGF(2alpha), and thromboxane B(2) (TxB(2)) were measured in EBC in 15 healthy ex-smokers, 20 steroid naïve patients with COPD who were ex-smokers, and in 25 patients with COPD who were ex-smokers and who were treated with inhaled corticosteroids. The study was of cross sectional design and all subjects were matched for age and smoking habit.

RESULTS

LTB(4) and PGE(2) concentrations were increased in steroid naïve (LTB(4): median 100.6 (range 73.5-145.0) pg/ml, p<0.001; PGE(2): 98.0 (range 57.0-128.4) pg/ml, p<0.001) and steroid treated patients with COPD (LTB(4): 99.0 (range 57.9-170.5) pg/ml, p<0.001; PGE(2): 93.6 (range 52.8-157.0) pg/ml, p<0.001) compared with control subjects (LTB(4): 38.1 (range 31.2-53.6) pg/ml; PGE(2): 44.3 (range 30.2-52.1) pg/ml). Both groups of patients had similar concentrations of exhaled LTB(4) (p=0.43) and PGE(2) (p=0.59). When measurable, LTE(4) and PGD(2)-methoxime concentrations were similar in COPD patients and controls, whereas PGF(2alpha) concentrations were increased in the former. TxB(2)-LI was undetectable in any of the subjects.

CONCLUSIONS

There is a selective increase in exhaled LTB(4) and PGE(2) in patients with COPD which may be relatively resistant to inhaled corticosteroid therapy.

Locally projecting GABAergic interneurons are the major providers of inhibition in the neocortex and play a crucial role in several brain functions. Neocortical interneurons are connected via electrical and chemical synapses that may be crucial in modulating complex network oscillations. We investigated the properties of spontaneous and evoked IPSCs in two morphologically and physiologically identified interneuron subtypes, the fast-spiking (FS) and low threshold-spiking (LTS) cells in layer V of rodent sensorimotor cortex. We found that IPSCs recorded in FS cells were several orders of magnitude more frequent, larger in amplitude, and had faster kinetics than IPSCs recorded in LTS cells. GABA(A) receptor alpha- and beta-subunit selective modulators, zolpidem and loreclezole, had different effects on IPSCs in FS and LTS interneurons, suggesting differential expression of GABA(A) receptor subunit subtypes. These pharmacological data indicated that the alpha1 subunit subtype is poorly expressed by LTS cells but makes a large contribution to GABA(A) receptors on FS cells. This was confirmed by experiments performed in genetically modified mice in which the alpha1 subunit had been made insensitive to benzodiazepine-like agonists. These results suggest that differences in IPSC waveform are likely attributable to distinctive expression of GABA(A) receptor subunits in FS and LTS cells. The particular properties of GABAergic input on different interneuronal subtypes might have important consequences for generation and pacing of cortical rhythms underlying several brain functions. Moreover, selective pharmacological manipulation of distinct inhibitory circuits might allow regulation of pyramidal cell activities under specific physiological and pathophysiological conditions.

BACKGROUND

Mosquito sampling methods are essential for monitoring and evaluating malaria vector control interventions. In urban Dar es Salaam, human landing catch (HLC) is the only method sufficiently sensitive for monitoring malaria-transmitting Anopheles. HLC is labour intensive, cumbersome, hazardous, and requires such intense supervision that is difficulty to sustain on large scales.

METHODS

Novel tent traps were developed as alternatives to HLC. The Furvela tent, designed in Mozambique, incorporates a CDC Light trap (LT) components, while two others from Ifakara, Tanzania (designs A and B) require no electricity or moving parts. Their sensitivity for sampling malaria vectors was compared with LT and HLC over a wide range of vector abundances in rural and urban settings in Tanzania, with endophagic and exophagic populations, respectively, using randomised Latin-square and cross- over experimental designs.

RESULTS

The sensitivity of LTs was greater than HLC while the opposite was true of Ifakara tent traps (crude mean catch of An. gambiae sensu lato relative to HLC = 0.28, 0.65 and 1.30 for designs A, B and LT in a rural setting and 0.32 for design B in an urban setting). However, Ifakara B catches correlated far better to HLC (r2 = 0.73, P < 0.001) than any other method tested (r2 = 0.04, P = 0.426 and r2 = 0.19, P = 0.006 for Ifakara A and LTs respectively). Only Ifakara B in a rural setting with high vector density exhibited constant sampling efficiency relative to HLC. The relative sensitivity of Ifakara B increased as vector densities decreased in the urban setting and exceeded that of HLC at the lowest densities. None of the tent traps differed from HLC in terms of the proportions of parous mosquitoes (P>>or= 0.849) or An. gambiae s.l. sibling species (P>>or= 0.280) they sampled but both Ifakara A and B designs failed to reduce the proportion of blood-fed mosquitoes caught (Odds ratio [95% Confidence Interval] = 1.6 [1.2, 2.1] and 1.0 [0.8, 1.2], P = 0.002 and 0.998, respectively), probably because of operator exposure while emptying the trap each morning.

CONCLUSIONS

The Ifakara B trap may have potential for monitoring and evaluating a variety of endophagic and exophagic Afrotropical malaria vectors, particularly at low but epidemiologically relevant population densities. However, operator exposure to mosquito bites remains a concern so additional modifications or protective measures will be required before this design can be considered for widespread, routine use.

Hyperlipidemia, one of the most important risk factors for coronary heart disease, is often associated with inflammation. We identified lymphotoxin (LT) and LIGHT, tumor necrosis factor cytokine family members that are primarily expressed on lymphocytes, as critical regulators of key enzymes that control lipid metabolism. Dysregulation of LIGHT expression on T cells resulted in hypertriglyceridemia and hypercholesterolemia. In low-density lipoprotein receptor-deficient mice, which lack the ability to control lipid levels in the blood, inhibition of LT and LIGHT signaling with a soluble lymphotoxin beta receptor decoy protein attenuated the dyslipidemia. These results suggest that the immune system directly influences lipid metabolism and that LT modulating agents may represent a novel therapeutic route for the treatment of dyslipidemia.

Blood lactate concentration ([La(-)](b)) is one of the most often measured parameters during clinical exercise testing as well as during performance testing of athletes. While an elevated [La(-)](b) may be indicative of ischemia or hypoxemia, it may also be a "normal" physiological response to exertion. In response to "all-out" maximal exertion lasting 30-120 seconds, peak [La(-)](b) values of approximately 15-25 mM may be observed 3-8 minutes postexercise. In response to progressive, incremental exercise, [La(-)](b) increases gradually at first and then more rapidly as the exercise becomes more intense. The work rate beyond which [La(-)](b) increases exponentially [the lactate threshold (LT)] is a better predictor of performance than V O2max and is a better indicator of exercise intensity than heart rate; thus LT (and other valid methods of describing this curvilinear [La(-)](b) response with a single point) is useful in prescribing exercise intensities for most diseased and nondiseased patients alike. H(+)-monocarboxylate cotransporters provide the primary of three routes by which La(-) transport proceeds across the sarcolemma and red blood cell membrane. At rest and during most exercise conditions, whole blood [La(-)] values are on average 70% of the corresponding plasma [La(-)] values; thus when analyzing [La(-)](b'), care should be taken to both (1) validate the [La(-)](b)-measuring instrument with the criterion/reference enzymatic method and (2) interpret the results correctly based on what is being measured (plasma or whole blood). Overall, it is advantageous for clinicians to have a thorough understanding of [La(-)](b) responses, blood La(-) transport and distribution, and [La(-)](b) analysis.

In order to definitively ascertain the functional contribution of lymphotoxin (LT) expressed by B cells, we produced mice with the LTbeta gene deleted from B cells (B-LTbeta KO mice). In contrast to systemic LTbeta deletion, in B-LTbeta KO mice only splenic microarchitecture was affected, while lymph nodes and Peyer's patches (PP) were normal, except for PP's reduced size. Even though B-LTbeta KO spleens retained a small number of follicular dendritic cells (FDC) which appeared to be dependent on LTbeta produced by T cells, IgG responses to sheep red blood cells were markedly reduced. Thus, the organogenic function of B-LTbeta is almost entirely restricted to spleen, where it supports the correct lymphoid architecture that is critical for an effective humoral immune response.